Variation of prolactin and β-Lactoglobulin genes in the Indonesian FH Cattle

Abstract Prolactin is a polypeptide hormone, encoded by the prolactin (PRL) gene, synthesized and secreted by anterior pituitary, and affecting milk yield and composition. β-Lactoglobulin (BLG) is the major whey proteinin the milk of ruminants. This study was conducted to identify the PRL and LGB genes polymorphism in the Indonesian FH cattle. A total of 139 individual cattle blood samples from West Java were used to obtain DNA samples through the DNA extraction process. Identification of the PRL and LGB genes was performed using PCR-RFLP method with RsaI (PRL gene) and HaeIII (BLG gene) restriction enzymes. The PRL gene was amplified using forward primer 5’-ccaaatccactgaattatgctt-3’ and reverse primer 5’-acagaaatcacctctctcattca-3’. The BLG gene was amplified using forward primer 5’-tgtgctggacaccgactacaaaaag-3’ and reverse primer 5’-gctcccggtatatgaccaccctct-3’. The PRL and BLG genes in the Indonesia FH cattle were polymorphic based on the PCR-RFLP analysis but the heterozygosity value was low. There were two alleles (G and A) and three genotypes (GG, GA, and AA) identified in the PRL gene of the Indonesian FH cattle with genotype frequencies were 0.914, 0.079, and 0,007 for GG, GA, and AA genotypes respectively. There were two genotypes (CC and CG) identified in the BLG gene with genotype frequencies were 0.91 (CC), and 0.09 (CG). Information about the PRL and BLG genes polymorphism in this study can be considered for further study to analyse its association with milk yield trait.

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Study of the Association between microRNA (miR-25T>C, miR-32C>A, miR-125C>T, and miR-222G>T) Polymorphisms and the Risk of Recurrent Pregnancy Loss in Korean Women

Genes ◽

10.3390/genes11040354 ◽

2020 ◽

Vol 11 (4) ◽

pp. 354 ◽

Cited By ~ 2

Author(s):

Jeong Yong Lee ◽

Jung Oh Kim ◽

Han Sung Park ◽

Chang Soo Ryu ◽

Ji Hyang Kim ◽

...

Keyword(s):

Pregnancy Loss ◽

Recurrent Pregnancy Loss ◽

Rflp Analysis ◽

Blood Coagulation Disorders ◽

Single Nucleotide Variants ◽

Genotype Combination ◽

Nitrogen Levels ◽

Genotype Frequencies ◽

Pcr Rflp ◽

Immunological Factors

Recurrent pregnancy loss (RPL), which is defined as two pregnancy losses that occur before 20 weeks of gestation, is relatively common, occurring in approximately 1–5% of women. The underlying cause is often unclear, although numerous factors may contribute to RPL, including environmental and immunological factors, blood coagulation disorders, and genetics. In particular, single nucleotide variants have been associated with RPL, including those found in microRNAs (miRNAs). We investigated the association between four miRNA polymorphisms, miR-25T>C, miR-32C>A, miR-125aC>T, and miR-222G>T, and RPL in a cohort consisting of 361 RPL patients and 272 controls. Subjects were genotyped at miRNA loci by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, and genotype frequencies were calculated. We then performed allele and genotype combination analyses and measured the association between miRNA polymorphisms and clinical variables in both RPL patients and controls. We detected a statistically significant association between RPL and the miR-25T/miR-32C/miR-125aT/miR-222T allele combination (adjusted odds ratio (AOR), 4.361; 95% confidence interval (CI), 1.496–12.72; P = 0.003). Three-gene combinations, including miR-32C/miR-125aT/miR-222T (AOR, 3.085; 95% CI, 1.254–7.588; P = 0.010) and miR-25T/miR-125aT/miR-222T (AOR, 2.929; 95% CI, 1.183–7.257; P = 0.015), and the two-gene combination miR-125aT/miR-222T (AOR, 2.417; 95% CI, 1.084–5.386; P = 0.026) were also associated with RPL. Analysis of variance (ANOVA) revealed that platelet counts and blood urea nitrogen levels were significantly different in RPL patients expressing different miR-125aC>T and miR-25T>C genotypes, respectively (P < 0.05). In addition, creatinine levels were lower in RPL patients expressing the minor alleles miR-25T>C and miR-32C>A. We investigated miRNAs (miR-25, miR-32, miR-125a, miR-222) in RPL patients and healthy controls. Significantly different allele frequencies were detected by ANOVA. We suggest that miRNAs and clinical factors can impact RPL occurrence.

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Identification of Pneumococcal Serotypes by PCR–Restriction Fragment Length Polymorphism

Diagnostics ◽

10.3390/diagnostics9040196 ◽

2019 ◽

Vol 9 (4) ◽

pp. 196 ◽

Cited By ~ 1

Author(s):

García-Suárez ◽

González-Rodríguez ◽

Cima-Cabal ◽

Yuste ◽

Vazquez ◽

...

Keyword(s):

Restriction Fragment Length Polymorphism ◽

Restriction Fragment ◽

Length Polymorphism ◽

Dna Sequences ◽

Fragment Length ◽

Fragment Length Polymorphism ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Pcr Rflp ◽

Restriction Fragment Length

Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dexB and aliA genes were compiled, and a computer screening of restriction enzymes was performed. The wzg–wzh–wzd–wze region and Sse9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates.

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PCR-based genetic identification of marlin and other billfish

Marine and Freshwater Research ◽

10.1071/mf98007 ◽

1998 ◽

Vol 49 (5) ◽

pp. 383 ◽

Cited By ~ 8

Author(s):

B. H. Innes ◽

P. M. Grewe ◽

R. D. Ward

Keyword(s):

Mitochondrial Dna ◽

Genetic Test ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Genetic Identification ◽

Dna Molecule ◽

Pcr Rflp ◽

D Loop ◽

Specific Variation ◽

Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

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Studies on TLR2 gene variants and their association with milk yield and milk quality traits in Bos indicus (Deoni) cattle

Indian Journal of Animal Research ◽

10.18805/ijar.b-3204 ◽

2017 ◽

Author(s):

U. T. Mundhe ◽

D. N. Das ◽

R. S. Gandhi ◽

P. Divya

Keyword(s):

Milk Yield ◽

Bos Indicus ◽

Restriction Enzymes ◽

Milk Quality ◽

Quality Traits ◽

Exon 2 ◽

Allelic Frequencies ◽

Tlr2 Gene ◽

Pcr Rflp

Present study molecular characterization of exon 2 of TLR2 gene and its association with milk yield and milk quality traits in 104 Deoni cattle using PCR- RFLP technique was done. Polymorphism was observed through HaeIII, HhaI and EcoRV restriction enzymes in Created Restriction Site (CRS) exon 2-1, CRS exon 2-5 and exon 2-1 by PCR- RFLP, respectively. In CRS exon 2-1 allelic frequencies were observed as 0.793 for A and 0.206 for B alleles and that of genotypic frequencies were 0.58 and 0.41 for genotypes AA and AB. In CRS exon 2-5, two genotypes viz., AC and CC with corresponding allelic frequencies were observed as 0.221 for A and 0.778 for C allele and that of genotypic frequencies observed were 0.44 and 0.55 for AC and CC genotypes respectively. TLR2 exon 2-1 exhibited two alleles G and T with frequencies of 0.134 and 0.865 and their Corresponding genotypic frequencies were 0.009, 0.25 and 0.74for GG, GT and TT genotypes respectively. Higher count of somatic cells (SCC) in TT homozygous and TG heterozygous genotypes, and lower in GG homozygous genotypes were observed in exon 2-1. Strongly significant (P£0.01) effect for least squares means of Test Day milk yield (TDMY) and Somatic Cell Count of CRS exon 2-1 were observed.

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The −174G/C and −572G/C Interleukin 6 Promoter Gene Polymorphisms in Mexican Patients with Rheumatoid Arthritis: A Case-Control Study

Clinical and Developmental Immunology ◽

10.1155/2013/959084 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 10

Author(s):

S. A. Zavaleta-Muñiz ◽

B. T. Martín-Márquez ◽

L. Gonzalez-Lopez ◽

N. G. Gonzalez-Montoya ◽

M. L. Díaz-Toscano ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Severe Disease ◽

Rflp Analysis ◽

Mexican Mestizo ◽

Lack Of Information ◽

Genotype Frequencies ◽

Pcr Rflp ◽

Promoter Gene ◽

Hardy Weinberg Equilibrium ◽

Control Study

Objective. There is a lack of information about the genotype frequencies of IL-6 −174G/C and −572G/C polymorphisms in Mexicans with rheumatoid arthritis (RA). Therefore, the aim of this study was to evaluate the association of the IL-6 −174G/C and −572G/C polymorphisms in Mexican mestizo with RA.Methods. We included 137 patients with RA and 102 healthy controls. Patients were assessed for clinical characteristics. IL-6 −174G/C and −572G/C polymorphisms were genotyped using PCR-RFLP analysis. Allele and genotype frequencies and the Hardy-Weinberg equilibrium were computed. Odds ratios (ORs) were computed to identify the risk for RA associated with the presence of GG genotype in comparison with the GC or CC genotypes.Results. The genotype −174GG occurred at a higher frequency in cases and controls (77.4% versus 78.4%,P=0.845). We found similar results for the genotype −572GG (54% in patients versus 60.8% in controls,P=0.295).Conclusions. This is the first study to evaluate the association of −174G/C and −572G/C polymorphisms of the IL-6 gene with RA in Mexican mestizo patients. These two polymorphisms were not associated with RA in the studied sample. Additional studies are required to evaluate if these IL-6 polymorphisms have relevance to the development of more severe disease.

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Effect of β-lactoglobulin gene polymorphism, lactation stage and breed on milk traits in Chios and Karagouniko sheep breeds

Annals of Animal Science ◽

10.1515/aoas-2016-0058 ◽

2017 ◽

Vol 17 (2) ◽

pp. 371-384 ◽

Cited By ~ 2

Author(s):

Kostas A. Triantaphyllopoulos ◽

Panagiota Koutsouli ◽

Athanassios Kandris ◽

Dimitris Papachristou ◽

Kalliopi E. Markopoulou ◽

...

Keyword(s):

Milk Yield ◽

Milk Traits ◽

Weinberg Equilibrium ◽

Sheep Breeds ◽

Lactation Stage ◽

Pcr Rflp ◽

Hardy Weinberg Equilibrium ◽

Daily Milk Yield ◽

Β Lactoglobulin ◽

Daily Milk

Abstract The animal selection with favourable phenotypes of the past has been, currently, replaced by the genotype selection on quantitative traits, assisted by the expanding molecular techniques in the context of livestock improvement. In this study, the c.112T>C polymorphism in exon II of β-lactoglobulin (β-LG) gene was investigated in Karagouniko and Chios sheep breeds by using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP), and possible associations with milk traits were examined. In total, 125 blood DNA samples were isolated for PCR-RFLP analysis and the respective 217 milk samples′ composition profile was obtained. The goodness of fit test to Hardy-Weinberg equilibrium (HWE) for β-LG genotypes was estimated and associations found between β-LG genotypes and raw milk composition. Two alleles and three genotypes were observed (AA, AB and BB) in both breeds, and Chios breed significantly deviated (P≤0.05) from Hardy-Weinberg equilibrium (HWE). Conclusively, linear mixed model analysis on samples, from both breeds collectively, showed significant effects of β-LG genotype on lactose percentage and somatic cell count (SCC), lactation stage on daily milk yield and protein, while the breed effect was significant only on daily milk yield.

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PCR–RFLP analysis of mitochondrial DNA for identification of Rutilus rutilus caspicus populations on the southern coast of the Caspian Sea, Iran

Journal of the Marine Biological Association of the United Kingdom ◽

10.1017/s0025315406014524 ◽

2006 ◽

Vol 86 (6) ◽

pp. 1463-1467 ◽

Cited By ~ 1

Author(s):

Sohrab Rezvani ◽

Amin Eimanifar ◽

Reza Aghili ◽

Faramarz Laloei

Keyword(s):

Caspian Sea ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Rutilus Rutilus ◽

Southern Coast ◽

The Caspian Sea ◽

Length Polymorphisms ◽

Pcr Rflp ◽

Nucleotide Divergence ◽

Genetic Pools

Genetic analysis using restriction fragment length polymorphisms (RFLPs) of cytochrome b in mtDNA was made to clarify genetic variations among two Iranian Rutilus rutilus caspicus populations of commercial importance from the southern coast of the Caspian Sea. Polymorphism was detected using six restriction enzymes and a total of six composite haplotypes were identified. Four haplotypes were rare occurring only once in two regions (west and east of the southern Caspian Sea). Nucleotide and haplotype diversities were higher in the south-west region of the Caspian Sea (π=3.43%, h=23.3).The nucleotide divergence between the two populations was low (0.064%). The test for heterogeneity of composite haplotype frequencies gave no significant outcome for all samples (χ2=0.137, P≤0.05). The results indicate that significant attention should be paid to the genetic characterization of R. rutilus caspicus populations for conservation of their genetic pools and aquaculture policies at the coastlines of the Caspian Sea.

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Detection and characterization of Leptospira spp. isolated from aborted bovine clinical samples

Acta Veterinaria Brno ◽

10.2754/avb201281010021 ◽

2011 ◽

Vol 81 (1) ◽

pp. 21-25 ◽

Cited By ~ 1

Author(s):

Hassan Momtaz ◽

Saadat Moshkelani

Keyword(s):

Public Health Problem ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Clinical Samples ◽

Dairy Herds ◽

Important Public Health Problem ◽

Pcr Rflp ◽

Beef Herds ◽

Bovine Abortion

Leptospira is recognized as an important public health problem worldwide, especially in tropical countries, and is a common cause of abortion in dairy and beef herds. The aim of the present study was to detect and characterize Leptospira as the causative agent of abortion in cattle using a PCR-RFLP in Chaharmahal va Bakhtiari and Isfahan provinces, Iran. A total of 220 bovine aborted foetuses and 120 vaginal discharges from an aborted calf were collected from 64 commercial dairy herds. After isolation of 60 Leptospira spp. from samples, RFLP analysis was carried out with HindIII and HaeIII restriction enzymes in reference strains and isolated for characterization. In a total of 340 specimens, 46 (20.9%) and 14 (11.66%) were identified positive for Leptospira spp. from aborted bovine foetuses and vaginal discharges, respectively. The present results also suggest that L. interrogans serovar hardjo has the highest prevalence in the region under study and L. hardjo is a major pathogen causing bovine abortion in Chaharmahal va Bakhtiari and Isfahan provinces of Iran.

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Genetic Variants of k-Casein and β-Lactoglobulin Genes and Their Association with Protein and Milk Components of Holstein Friesian Cows at Small Farmers in Lembang, West Java

KnE Life Sciences ◽

10.18502/kls.v2i6.1023 ◽

2017 ◽

Vol 2 (6) ◽

pp. 86 ◽

Cited By ~ 1

Author(s):

Anggraeni Anggraeni ◽

A. Anneke ◽

H.S. Nury ◽

E. Andreas ◽

C. Sumantri

Keyword(s):

Genetic Variants ◽

Milk Protein ◽

Restriction Enzymes ◽

Dairy Farmers ◽

West Java ◽

Holstein Friesian ◽

Pcr Rflp ◽

Milk Components ◽

Milk Component ◽

Β Lactoglobulin

Genetic variants of CSN3 and LGB genes and their effects on protein and milk components were studied in Holstein Friesian at small dairy farmers in Lembang District, West Java, Indonesia. Allelic variants were identified by PCR-RFLP technique using restriction enzymes of Pst I for the CSN3 gene and Hae III for the LGB gene. The CSN3 gene was dominated by AB genotype. Milk protein was not affected by genotypes of the two genes. Only fat content was significantly affected (P <0.05) by the CSN3 gene with AB cows having the highest fat to AA and BB cows (3.76% vs. 3.26% and 3.34%). Keywords: CSN3 gene; LGB gene; milk protein; and milk component.

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PHYLOGENETIC RELATIONSHIPS AMONGST 10 Durio SPECIES BASED ON PCR-RFLP ANALYSIS OF TWO CHLOROPLAST GENES

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v6n1.2005.20-27 ◽

2013 ◽

Vol 6 (1) ◽

pp. 20 ◽

Cited By ~ 2

Author(s):

Panca J. Santoso ◽

Ghizan B. Saleh ◽

Norihan M. Saleh ◽

Suhaimi Napis

Keyword(s):

Phylogenetic Analysis ◽

Phylogenetic Relationships ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Rbcl Gene ◽

Research Station ◽

Taxonomic Level ◽

Pcr Rflp ◽

Young Leaves ◽

Reliable Marker

Twenty seven species of Durio have been identified in Sabah and Sarawak, Malaysia, but their relationships have not been studied. This study was conducted to analyse phylogenetic relationships amongst 10 Durio species in Malaysia using PCR-RFLP on two chloroplast DNA genes, i.e. ndhC-trnV and rbcL. DNAs were extracted from young leaves of 11 accessions from 10 Durio species collected from the Tenom Agriculture Research Station, Sabah, and University Agriculture Park, Universiti Putra Malaysia. Two pairs of oligonucleotide primers, N1-N2 and rbcL1-rbcL2, were used to flank the target regions ndhC-trnV and rbcL. Eight restriction enzymes, HindIII, BsuRI, PstI, TaqI, MspI, SmaI, BshNI, and EcoR130I, were used to digest the amplicons. Based on the results of PCR-RFLP on ndhC-trnV gene, the 10 Durio species were grouped into five distinct clusters, and the accessions generally showed high variations. However, based on the results of PCR-RFLP on the rbcL gene, the species were grouped into three distinct clusters, and generally showed low variations. This means that ndhC-trnV gene is more reliable for phylogenetic analysis in lower taxonomic level of Durio species or for diversity analysis, while rbcL gene is reliable marker for phylogenetic analysis at higher taxonomic level. PCR-RFLP on the ndhC-trnV and rbcL genes could therefore be considered as useful markers to phylogenetic analysis amongst Durio species. These finding might be used for further molecular marker assisted in Durio breeding program.

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