Genetic structure features of cattle populations of Ukrainian selection by polymorphism of loci, associated with milk productivity traits

2019 ◽  
Vol 6 (3) ◽  
pp. 37-44
Author(s):  
R. Kulibaba ◽  
Yu. Liashenko ◽  
P. Yurko
Keyword(s):  
Genetic Structure ◽  
Restriction Analysis ◽  
Placental Lactogen ◽  
Functional Genes ◽  
Black And White ◽  
Prolactin Gene ◽  
Rsai Polymorphism ◽  
Experimental Populations ◽  
Dairy Breed ◽  
Fourth Exon

Aim. To study the genetic structure of cattle populations of Ukrainian selection by polymorphism of functional genes (PRL, PL) and microsatellites (BM027, RM185). Methods. The study was conducted using the method of polymerase chain reaction (PCR) and restriction analysis in case of loci PRL and PL, and using classic PCR with subsequent electrophoresis in polyacrylamide gel to analyze microsatellite variability. Results. The results of the studies demonstrated that the locus of prolactin by RsaI-polymorphism in the fourth exon was polymorphic in both experimental populations (Ukrainian Black-and-White and Ukrainian Red-and-White dairy breeds of cattle). The mutation (Indel) was fi rst determined in the fourth exon of prolactin gene, the variants of which correlated with some alleles of the locus by RsaI-polymorphism. The locus of placental lactogen by RsaI-polymorphism in the fi fth exon was monomorphic in both experimental populations. Microsatellite locus RM185 was polymorphic in both groups of animals, whereas BM027 – only in the Black-and-White dairy breed. Conclusions. The specifi cities of the genetic structure of the Ukrainian Black-and-White and Red-andWhite dairy breed populations by polymorphism of functional genes and microsatellite loci were determined. The locus of placental lactogen by RsaI-polymorphism in the fi fth exon cannot be used in further studies due to the absence of alternative variants of the gene in both studied populations of animals. The analysis of the distribution of haplotype frequencies demonstrated the absence of deviation from the equilibrium state by linkage for each of the investigated markers which makes their use impossible in the breeding programs as a separate functional unit.

scholarly journals POLYMORPHISM OF GROWTH HORMONE RECEPTOR GENE AND MYOGENIC FACTOR 5 GENE IN DAIRY CATTLE POPULATIONS

2021 ◽  
pp. 69-78
Author(s):  
H. S. Alshamaileh ◽  
R. O. Kulibaba ◽  
Yu. Liashenko ◽  
H. S. Borzova
Keyword(s):  
Growth Hormone ◽  
Genetic Structure ◽  
Equilibrium State ◽  
Allele Frequency ◽  
Hormone Receptor ◽  
Growth Hormone Receptor ◽  
Restriction Analysis ◽  
Black And White ◽  
Myogenic Factor ◽  
Dairy Breed

Investigations of the genetic structure features of the populations of Ukrainian Black-and-White and Red-and-White dairy breeds by polymorphism of growth hormone receptor (GHR) and myogenic factor 5 (MYF5) genes were carried out. AluI-polymorphism in the promoter fragment of the GHR gene and TaqI-polymorphism in the second intron of the MYF5 gene was detected using polymerase chain reaction and restriction analysis (PCR-RFLP). According to the research results, it was shown that the growth hormone receptor and myogenic factor 5 genes in the populations of Ukrainian Black-and-White and Red-and-White dairy breeds are polymorphic. In the population of the Ukrainian Black-and-White dairy breed at the GHR locus, the frequency of the AluI+ allele was 0.61; allele AluI- – 0.39; at the MYF5 locus, the TaqI+ allele frequency was 0.65; allele Taq- – 0.35, respectively. In the population of the Ukrainian Red-and-White dairy breed at the GHR locus, the frequency of the AluI+ allele was 0.54; allele AluI- – 0.46; at the MYF5 locus, the TaqI+ allele frequency was 0.64; allele Taq- – 0.36, respectively. Both experimental populations for GHR and MYF5 loci are in a genetic equilibrium state according to Hardy-Weinberg, which indicates the absence of microevolutionary changes in the process of their reproduction. There were no significant fluctuations in the genetic structure according to the revealed features of the genotype and allele frequencies distribution. The breeding work that is carried out on both populations does not affect marker alleles (which are described in the work), which is reflected in the features of the genetic-population structure of the experimental groups and their equilibrium state. The features of the distribution of individuals with different genotypes by GHR and MYF5 loci in the populations of both breeds make it possible to carry out further studies to determine the relationship between the identified allelic variants of polymorphic loci with parameters of animal productivity.

scholarly journals Association of genetic variants of bovine <i>prolactin</i> with milk production traits of Black-and-White and Jersey cattle

2005 ◽  
Vol 48 (2) ◽  
pp. 149-156 ◽  
Author(s):  
A. Dybus ◽  
W. Grzesiak ◽  
H. Kamieniecki ◽  
I. Szatkowska ◽  
Z. Sobek ◽  
...  
Keyword(s):  
Milk Production ◽  
Production Traits ◽  
Milk Production Traits ◽  
Black And White ◽  
Jersey Cattle ◽  
Prolactin Gene ◽  
Pcr Rflp ◽  
Rsai Polymorphism ◽  
White Cattle ◽  
Jersey Cows

Abstract. Associations between polymorphism localised in the third exon of the prolactin gene (PRL-RsaI) and milk xproduction traits of Black-and-White and Jersey cattle were analysed. A total of 427 cows were included in the study. PCR-RFLP method was used. The frequencies of genotypes and alleles were as follows: 0.7107 – AA, 0.2851 – AB, 0.0042 – BB; 0.8533 – PRLA and 0.1467 – PRLB for Black-and-White cattle and 0.0919 – AA, 0.4324 – AB, 0.4757 – BB; 0.3081 – PRLA and 0.6919 – PRLB for Jersey cattle. Statistically significant differences between the breeds were observed in the frequencies of genotypes and alleles. Associations between PRL-RsaI polymorphism and milk production traits of Jersey cows and lack of associations with these traits in Black-and-White cows were observed.

scholarly journals Polymorphism of LEP and TNF-α Genes in the Dairy Cattle Populations of Ukrainian Selection

2021 ◽  
Vol 34 (1) ◽  
pp. 180-191
Author(s):  
Roman Kulibaba ◽  
Yuriy Liashenko ◽  
Mykola Sakhatskyi ◽  
Yulia Osadcha ◽  
Hamza Alshamaileh
Keyword(s):  
Dairy Cattle ◽  
Restriction Analysis ◽  
Low Frequency ◽  
Cattle Breed ◽  
Fixation Index ◽  
Necrosis Factor Alpha ◽  
Black And White ◽  
Tnf Α ◽  
Hardy Weinberg Equilibrium ◽  
Dairy Breed

Study was conducted by investigating the genetic structure of cattle populations of Ukrainian Black-and-White and Ukrainian Red-and-White dairy breeds by polymorphism of leptin gene (LEP) and tumor necrosis factor-alpha gene (TNF-α). The studies were carried out using the method of polymerase chain reaction (PCR) and restriction analysis for LEP and using classic PCR with subsequent SSCP analysis in case of TNF-α. According to the results of the study, it was shown that the LEP gene was polymorphic in both experimental populations by HphI-polymorphism in the third exon of the gene (A59V mutation). In the population of the Ukrainian Black-and-White dairy cattle breed, the frequency of allele C (HphI-) was 0.77; allele T (HphI+) – 0.23. Wright's fixation index was 0.23; which indicates a significant excess in the number of homozygous individuals. This population had a deviation from the Hardy-Weinberg equilibrium state. In the population of the Ukrainian Red-and-White dairy breed, the frequency of allele C was 0.72; allele T – 0.28; Wright's fixation index was -0.18. According to the SSCP-polymorphism of the TNF-α gene, 6 alleles with a size of 450-1200 bp were detected (alleles A, B, and F for Ukrainian Black-and-White dairy breed; A, B, C, D, Е, F – for Ukrainian Red-and-White dairy breed). The frequency of allele A prevailed in the populations of both breeds (0.58 and 0.54, respectively). Alleles C, D, and E had a low frequency of occurrence (0.04-0.16) and they were found only in the population of Red-and-White dairy breed.

Linear type trait evaluation of firstborns of Ukrainian Black-and-White Dairy breed depending on the body build

2019 ◽  
pp. 24-34
Author(s):  
R. Stavetska ◽  
Y. Dynko ◽  
M. Bushtruk ◽  
I. Starostenko ◽  
O. Babenko ◽  
...  
Keyword(s):  
Body Condition Score ◽  
High Capacity ◽  
The Body ◽  
Body Depth ◽  
Linear Type ◽  
Body Build ◽  
Black And White ◽  
Condition Score ◽  
Type Traits ◽  
Dairy Breed

It has been experimentally proven that the strength and direction of the connection between the linear type traits evaluation of the Ukrainian black-and-white dairy breed cows and their body build depends on the type of the build and the sign of the exterior. The study was conducted in 2018 on a breeding plant of the Ukrainian Black-and-White Dairy breed SPC named Shchorsa in Kiev region (n=89). Differentiation of cows on types of their body build was conducted by the method proposed by Chernenko (low-capacity, mid-capacity and high-capacity types) and Kolesnik (by the massiveness index – on the friable and dense, the bone index – delicate and rough, the wide-breast and wide-rump indices – on narrow-body and wide-body types). It was found that the cow-heiferswere well-developed, had intermediate stature and optimum body condition score, deep body, and wide, slight slope rump. They alsohad desirable rear legs, set and foot angle but some toe-out, the udder was well-balanced. Under the good total score of the exterior of studied cows there were some animals with short stature (134– 136 cm), very wide rump, straight rear legs side view and toe-out, extremely steep foot angle, loose fore udder attachment, deep udder below hocks and front teat placement inside of quarter. The degree of development of the studied linear traits of cow-heifers confirms their in-herd variability. The variability of the exterior traits varies from 10,7 % (body depth) to 38,6 % (stature). In addition to the stature of a significant variability was characterized of angularity (22,4 %), rear legs set (22,5 %), body condition score (21,8 %), as well as traits that describe udder: rear udder attachment (31,4 %), udder depth (32,6 %) and front and rear teat position (31,3 % and 25,1 %, respectively). The high variability of linear traits indicates the possibility of effective their selection. As a result of the research, it has been established that the strength and direction of the correlations between the linear type traits evaluation of Ukrainian Black-and-White Dairy breed cows and the type of their body build depends on the type of the body build and the linear trait. The low-, mid- and high-capacity bodies build types (according to Chernenko) reliably correlate with chest width (r=0,58) and body depth (r=0,48), rear legs rear view (r=0,30), the central ligament (r=0,28), udder depth (r =-0,22) and teat length (r=0,25), P˂0,01–0,001. Friable and dense bodies build (according to Kolesnik) were most strongly associated with stature (r=-0,28), chest width (r =-0,70), and body depth (r=-0,38), as well as with rear legs rear view (r=-0,23), P˂0,05...0,001; delicate and rough bodies build – with stature (r=-0,51), angularity (r=-0,22),rump width (r=-0,35), udder depth (r=-0,27) and rear udder attachment (r =0,20), P ˂ 0,05–0,001; narrow-body and wide-body types – with chest width (r=0,30) and rump width (r=0,23), P˂0,05–0,01. Linear type trait evaluation of cows depends on the body build. The strength and direction of the correlations between types of body build and linear traits showed that the most informative is the classification of the body build by low-, mid- and high-capacity according to Chernenko – on the average η2x=17,5 %. The less informative – by narrow-body and wide-body types according to Kolesnik – η2x=7,6 %. The classification of cows on delicate-rough, narrow-body and wide-body types (according to Kolesnik) according to informativeness and reliability can have the intermediate position – η2x=10,0–11,5 %. The strongest influence on the body build has the cows’ height (η2x=27,9 % on average for all studied types of body build), chest width (η2x=33,5 %), body depth (η2x=21,8 %), rump width (η2x=24,1 %) and rear legs (η2x=18,7 %). Key words: cows, linear type trait evaluation, body build, correlation, power of influence.

CONSEQUENCES OF “HOLSTEINIZATION” OF UKRAINIAN BLACK-PIED DAIRY BREED BY GENE BOLA-DRB3.2

2017 ◽  
Vol 54 ◽  
pp. 146-156
Author(s):  
T. M. Suprovych ◽  
M. P. Suprovych ◽  
R. V. Kolinchuk
Keyword(s):  
Genetic Distance ◽  
Milk Production ◽  
Genetic Similarity ◽  
Restriction Analysis ◽  
Allelic Polymorphism ◽  
Milk Productivity ◽  
High Productivity ◽  
Black And White ◽  
Holstein Breed ◽  
Specific Pcr

Introduction. The main direction of increasing the productivity of milk is to increase the proportion of heredity of the Holstein breed in the genotype of cows. Industrial breeds in Ukraine are improving due to the increase in the Holstein inheritance in the genotype of cows. The "holsteinization" of the most widespread domestic Black-and-White diary breed is intensively conducted. Currently, the percentage of heredity from Holstein is 90% or more. The negative effect of "holsteinization" appeared in reducing the resistance of animals to diseases that led to the spread of necrobacterial pathology. The control of the spread of necrobacteriosis can be based on genetic markers. Important markers can be the allele of the BoLA-DRB3.2 gene responsible for the formation of adaptive immunity. Due to the ambiguity of the results of "holsteinization", the following tasks were solved: To study the genetic structure of the herd for the BoLA-DRB3.2 gene at the beginning of the "holsteinization" and now. To compare the detected genetic structures with the alleles spectrum of North American Holstein and identify quantitative and qualitative changes in the structure of the herd genotype. To determine the effect of "holsteinization" on the dynamics of milk production and the state of morbidity by necrobacteriosis. Materials and methods of research. Comparison of alleles of population of the Ukrainian Black-Pied Dairy (UBPD) breed and Holstein breed was conducted to detect the consequences of "holsteinization" on milk yield and incidence of necrobacteriosis. The data of the allelic polymorphism of the BoLA-DRB3.2 gene of the UBPD10 (2010, n = 162), UBPD15 (2015, n = 114) and two Holstein populations of the USA and Canada were collected. The allelic spectrum was determined by the PCR-RFLP method. The amplification of the BoLA-DRB3.2 gene was performed using 2-step PCR with the use of primers HLO-30, HLO-31 and HLO-32 and allele-specific PCR. Restriction analysis was performed with endonuclease RsaI, HaeIII, BstYI (XhoII). Restriction fragments were separated by electrophoresis in 4% agarose gel. Counting of allele frequencies was performed taking into account the number of homozygotes and heterozygotes found for the corresponding alleles. To determine the phylogenetic relationships between the populations of the studied herds, genetic distance and genetic similarity were determined by the M. Nei method. Individual dairy productivity of cows was estimated for all lactation (regardless of its duration). Average milk yields were determined as the total volume of milk produced divided by the number of dairy cows. Results and discussion. The breeding measures carried out led to the accumulation of alleles characteristic of the Holstein breed. For Holstein, there are eight alleles with a frequency of more than 4%. It is alleles *03, *07, *08, *11, *16, *22, *23, *24. A high degree of consolidation of weighty alleles can be outlined. In total they occupy 84,6% of allele spectrum of the population. Consolidation of such alleles in the herd of the Ukrainian Black-and-White diary breed is much lower - only 52.2%, although it increased by 6.2% over 5 years. Alleles *10, *13 and *28 are "weighty" for the Ukrainian Black-and-White diary breed, but they are almost non-existent in Holsteins. The genetic similarity of the herd UBPD15 and Holstein increased by ΔI = 0,085, and the genetic distance between the herds of the UBPD increased by ΔD = 0,085 for 5 years. The comparison of the allele spectrum of Holstein and the Ukrainian Black-and-White diary breed shows both the accumulation and the elimination of alleles associated with high productivity. The largest consolidation is typical for alleles *24 (+ 6.75%) and *16 (+ 4.65%). The frequency of "milk" alleles *22 and *08 decreased, respectively, by 4.14 and 1.27%. Alleys, which cause low milk productivity, have the following dynamics: * 23 + 2.53%, *11 – 0.67 and *28 – 0.26. The accumulation of alleles *16 and *23 (7.18%) was found that are associated with predisposition to necrobacteriosis and elimination of *03 and *22 alleles (4.75%) that influence on this disease. Conclusions. It is determined that the role of alleles characteristic for Holstein is increasing in the the Ukrainian Black-and-White diary herd. Breeding measures for holsteinization are conducted in the right direction. There is accumulation of alleles associated with high milk productivity and predisposition to necrobacteriosis. It positively affects the growth of milk production and negatively affects the incidence of necrobacteriosis.

FEATURES OF THE EXTERIOR TYPE OF UKRAINIAN BLACK-AND-WHITE DAIRY BREED OF THE CHERKASSY REGION, ESTIMATED BY THE LINEAR CLASSIFICATION METHOD

2017 ◽  
Vol 54 ◽  
pp. 112-119
Author(s):  
L. M. Khmelnychy
Keyword(s):  
Local Population ◽  
The Body ◽  
Breeding Value ◽  
Linear Classification ◽  
Body Structure ◽  
Linear Type ◽  
Maternal Line ◽  
Black And White ◽  
Type Traits ◽  
Dairy Breed

The estimation of the first-born cows of Ukrainian Black-and-White dairy breed by the exterior type in the leading breeding herds of Cherkassy region by the method of linear classification (n = 566) has been carried out. The results of the evaluated animals by the 100-score system indicate that within the group of traits, the average level of assessment is "good with the plus". The animals have been differed by the good development of group features characterizing the dairy type (82.4 score), body condition (83.5 score), limbs (82.8 score), udder (82.7 score) and overall score (82.8 score). The degree of development 18 descriptive traits of the exterior of cows, has been predicted by the method of linear classification, indicates their significant intraherd variability. Animals have the moderate height, deep body and angularity. The rump angle, the angle of hock joint and the teats length, according to the variability of scores in the range of 17.8–22.4%, have an average optimal development. The level of scores for the rear width (5.9 score), attachment of the fore (6.3 score) and the rear parts (6.0 score) of the udder testifies to their good development. The body structure of the cows-firstborn has a sufficient description of the type traits that determine their milk yield. At the present stage of selection and breeding work due to the use of bull-sires different origins and breeding value on the local population livestock with significant genotype diversity, the degree of influence each of the hereditary factors in the overall variability of linear features has a scientific and practical significance. In our studies, the force of influence conditional bloodiness of the Holstein mother's breed in the overall fraction of variability of linear type traits within the exterior complexes and the overall assessment takes a significant percentage - from 24.9 to 31.5 with a high degree of reliability according to Fischer's criterion. Among the descriptive traits, the proportion of influence the conditional bloodiness of mother, which is confirmed by high statistical confidence, ranges from 14.4 to 19.8%, and refers to the traits important for breeding. The level of coefficients of influence the father's breeding value in the total phenotypic variability for some of the linear traits is high and reliable. First of all it concerns group traits of the exterior, the development of which is determined by the pedigree value of father by 37.1–41.8%, and the overall assessment of type – by 46.6%. The coefficients of influence father's breeding value in most of the descriptive traits have high reliability (P < 0,001). The coefficients of the force of influence father, depending on the heredity of Holstein for the linear traits of almost the same level as his pedigree value. Since in this case the organized factor is purebreed Holstein and crossbreed bulls-sires of domestic selection, the positive role of Holstein heredity in improving the exterior type of cows has been traced realistically. The necessity of linear breeding, as an effective method in the system of dairy cattle breeding, has been substantiated by reliable values of the coefficients of the force of influence of father's line on the linear traits of offspring (3.4–29.5%). Despite the significant variability of coefficients of the force of influence on the linear traits, most of them have a high level of reliability within the evaluated body parts important for selection. The effect of cows belonging to the maternal line on the development of features of the body structure is insignificant and unreliable (5.3–9.2%). In the process of breeding aimed at improving cows of dairy herd by the exterior type, the pedigree value of parents has been assessed according to the method of linear classification of their daughters and their belonging to perspective genealogical formations.

INFLUENCE OF LIVE WEIGHT IN CULTIVATION ON THE DURATION OF PRODUCTIVE USE OF COWS OF UKRAINIAN BLACK-AND-WHITE DAIRY BREED OF DIFFERENT BLOODLINE ON THE HOLSTEIN BREED

2020 ◽  
Vol 11 (2) ◽  
pp. 16-27
Author(s):  
L. M. Danets ◽  
I. V. Tkachova ◽  
V. P. Shablia
Keyword(s):  
Average Duration ◽  
Live Weight ◽  
Black And White ◽  
Holstein Breed ◽  
Complex Integral ◽  
The Cost ◽  
State Enterprises ◽  
Dairy Breed ◽  
Integral Feature ◽  
A Share

The duration of productive use is a complex integral feature and is determined by both genetic and paratypical factors. This feature should ensure maximum milk productivity of animals, economic efficiency of dairy farms and generally limit the cost of raising and keeping cows. Research conducted in the experimental farms of state enterprises "Kutuzovka", Kharkov district of Kharkov region using cow’s Ukrainian black-and-white dairy breed (4038 cows). We studied the duration of productive use of cows divided into gradations according to the conditional bloodline for the Holstein breed, depending on the weight in the control age periods of cultivation (at birth, at 6, 12 and 18 months). The maximum value of the duration of productive use was recorded in the group of cows with a share of conditional blood for the Holstein breed up to 30% inclusive (the average duration of productive use is 2.77 lactations). The highest indicator of the duration of productive use of the studied animals was 4.09 lactations. Among cows with a share of conditional blood for the Holstein breed up to 30% longer produced those born with a weight of more than 40 kg (4.09 lactations), at 6 months of age weighed 100-149 kg, at 12 months – 200-249 kg, at 18 months of age – 350-399 kg. Among animals with a share of bloodline of 31-60 %, those that had a weight at the age of 6 months produced the longest: 155-190 kg (3.17 lactations), at 12 months – 250-299 kg (2.98 lactations), at 18 months – 350-399 kg (3.06 lactations). In the most numerous gradation with the share of bloodline for the Holstein breed 61-90 %, the longest productive use was characterized by animals born with a weight of 30-39 kg (2.12 lactation), at 6 months of age they weighed more than 200 kg (3.29 lactation), at 12 months-300-349 kg (3.40 lactation), at 18 months – 400-449 kg (2.82 lactation). The average duration of productive use of cows of this grade is quite low – in the range of 2.82-3.29 lactations. The highest degree of influence on the duration of productive use in this gradation of animals was recorded by the live weight factor at 6 months of age (η2 = 10.8). Сcomparative assessment of the cows gradation with a share of conditional blood for the Holstein breed of 91 % or more found that the longest productive use was characterized by cows that had a live weight at birth of 30-39 kg (1.93 lactation), in 6 months – 155-199 kg (2.73 lactation), in 12 months – 250-299 kg (2.87 lactation) and in 18 months – 350-399 kg (2.62 lactation). The degree of influence of live weight at this age on the duration of productive use of cows is insignificant, except for live weight at 6 months of age (η2 = 11.8).

Population characteristics of daughters of domestic and foreign breeding bulls

2021 ◽  
Vol 1 (2) ◽  
pp. 29-35
Author(s):  
N. I. Abramova ◽  
◽  
O. L. Khromova ◽  
M. O. Selimyan ◽  
◽  
...  
Keyword(s):  
Genetic Structure ◽  
Milk Yield ◽  
Population Characteristics ◽  
Breeding Value ◽  
Black And White ◽  
Vologda Region ◽  
Breeding Work ◽  
White Breed ◽  
Productive Traits ◽  
Modern Population

The use of foreign-bred bulls leads to changes in the genetic structure of the black-and-white breed population, which affects the level of population characteristics. The study was conducted to study the comparative variability and heritability of productive and reproductive characteristics of daughters of sires of domestic and foreign selection in pedigree livestock 10 250 cows 1st calving modern population of black-motley breed of the Vologda region. The superiority of the offspring of foreign bulls in the average indicators of all the studied traits was established with confidence (P<0,001). There were no significant differences in the level of variability of the characteristics of the descendants of domestic and foreign bulls. According to productive traits, the greatest variability of indicators was established for milk yield for 305 days of the 1st lactation Cv = 17,3 – 17,7%, therefore, selection on this trait will be more effective. The greatest share of the influence of bullsfathers, both domestic and foreign selection, in the total sum of all factors determining the value of traits in the population of the black-and-white breed, is determined by the milk yield of daughters η2 = 0,22 – 0,24, the age of the 1st fruitful insemination and the first calving η2 = 0,24 – 0,25. In further breeding work with the breed, it is necessary to obtain and use domestic producers that exceed the breeding value of foreign bulls.

scholarly journals FEATURES OF DIAGNOSIS OF NECROBACTERIOSIS OF COWS BY PCR-RFLP

2020 ◽  
pp. 26-37
Author(s):  
O.D. Biriukova ◽  
T.M. Suprovych ◽  
M.P. Suprovych ◽  
S.V. Laiter-Moskaliuk ◽  
I.O. Chornyi
Keyword(s):  
Molecular Genetic ◽  
Experimental Sample ◽  
Exon 2 ◽  
Blood Samples ◽  
Restriction Fragments ◽  
Black And White ◽  
Pcr Rflp ◽  
Dairy Breed ◽  
Established Technique ◽  
Gene Exon

Molecular genetic markers can detect polymorphism at the DNA level. This feature determines the possibility of their widespread use in genetics and breeding. Alleles of the BoLA-DRB3 gene (exon 2) can act as such markers if a statically significant association between the disease and the allele is established. The presence of such DNA markers in the genotype of animals makes it possible to judge the likelihood of disease in postnatal ontogenesis immediately after the birth of a heifer, based on which we can conclude about the conditions of further use of the animal in the main herd. According to the results of studying the polymorphism of the BoLA-DRB3 gene in cows of the Ukrainian black and white dairy breed resistant and susceptible to necrobacteriosis, four "informative" alleles were revealed. Two of them *03 and *22 are associated with resistance, and the other two - *16 and *23 with susceptibility to necrobacteriosis. The presence of these alleles in the genotype of the animal is determined by testing performed by PCR-RFLP. The method is time consuming, labor intensive and costly. To simplify it, the following technique is proposed. Restriction fragments of alleles *03, *16, *22 and *23 for endocluases RsaI, XhoII and HaeIII have the following DNA patterns: bbb, jbd, mba and nba. Due to the peculiarity of the restriction fragments, which is that endonuclease XhoII reveals in these alleles only one pattern b with length of 284 bp, the process of determining informative alleles can be simplified. Isolation of DNA from blood samples and amplification of a fragment of the BoLA-DRB3.2 gene with a size of 284 bp is carried out according to the established technique. Next, the restriction of the fragment by endonuclease XhoII and sampling having a pattern b. Selected samples are treated with RsaI endonuclease and only those with patterns b, j, m and n remain. The next step is to restrict the selected samples with HaeIII endonuclease and select heifers with bbb (*03) and nba (*23) genotypes. After the first restriction, blood samples without pattern b are eliminated from the experimental sample; after the second – two alleles with patterns RsaI + XhoII jb (*16) and mb (*22) are unambiguously determined, after the third – genotypes bbb and nba, which correspond to alleles *03 and *23. In total, only 75% of blood samples are typed, which reduces the material consumption, time and cost of work to identify heifers genetically susceptible (resistant) to necrobacteriosis.

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