​Identification of Single Nucleotide Polymorphism (SNP) C-2161G in the Promoter Region of Prolactin Gene and its Association with Egg Production in Tellicherry Native Chicken

Author(s):  
Azhaguraja Manoharan ◽  
S. Sankaralingam ◽  
P. Anitha ◽  
Binoj Chacko ◽  
T.V. Aravindakshan

Background: Prolactin is a physiological candidate gene which has significant effects on egg production in poultry. Also, it plays a major role on incubation behaviour (broodiness) in birds. The main objective of the present study was identification of single nucleotide polymorphism (SNP) C-2161G in the promoter region of prolactin gene and its association with egg production in Tellicherry native chicken population. Methods: A total of 200 blood samples were collected from the randomly selected birds of Tellicherry native chicken in All India Co-ordinated Research Project on Poultry improvement (AICRP) farm, Mannuthy, Thrissur, Kerala. Isolation of Genomic DNA was done and the isolated samples were subjected to polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis to identify the SNP C-2161G of prolactin gene. Result: On the basis of RFLP patterns, birds were designated with three different genotypes namely CC, CG and GG. The allelic and genotypic frequency was calculated. The observed genotypic frequency at the SNP site C-2161G was CC (0.61), CG (0.160) and GG (0.23) and the frequency of allele was 0.69 for C and 0.31 for G. The egg production was shown to be statistically similar for the genotypes of SNP C-2161G.

2018 ◽  
Vol 68 (3) ◽  
pp. 391 ◽  
Author(s):  
MOHAMED M.M. OSMAN ◽  
SHAABAN A. HEMEDA ◽  
ABEER A.I. HASSANIN ◽  
WALAA A. HUSSEINY

Broodiness is a behavioral trait observed in most common breeds of domestic fowl and due to its fundamental role in avian reproduction, it has been of great interest to poultry scientists, breeders and producers of hatching eggs. Prolactin gene (PRL) is generally accepted as crucial to the onset and maintenance of broodiness in birds and thus plays a crucial role in egg production. Therefore, the present study aimed to screen the Single Nucleotides Polymorphisms (SNPs) of prolactin gene in four commercial chicken lines namely Hubbard F15, Lohmann, Cobb500, and Avian48 using PCR and direct sequencing. A total number of forty chickens (ten females from each of the four commercial chicken lines) were used. Blood samples were collected aseptically from brachial (wing) vein of the chickens for genomic DNA extraction. PCR reaction was done using five pairs of primers, one sense (F) and one antisense (R) primer for each of the five exons of prolactin gene. Finally, DNA sequencing and Single Nucleotide Polymorphisms (SNPs) analysis was done using Laser gene Megalign program. The results showed three SNPs in Hubbard F15 chicken line; one synonymous SNP at the position 3838 bp (ACC/ACT-transition) in exon 2 while in exon 5, two SNPs were detected; one non-synonymous single nucleotide polymorphism at the position 7921bp (CCT/TCT-transition) which results in amino acid changes at codon positions 169 (P/S), and one synonymous single nucleotide polymorphism at the position 8187 bp T/ C. The study concluded that this SNP in PRL gene could be used as the potential molecular markers for egg production traits in chicken.


Author(s):  
Azhaguraja Manoharan ◽  
S. Sankaralingam ◽  
P. Anitha ◽  
Binoj Chacko ◽  
T. V. Aravindakshan

Background: Broodiness is a sex linked behavioural trait observed in most of the domestic fowls and it’s also known as incubation behaviour. Prolactin (PRL) is the principal gene which plays a crucial role to the onset and maintenance of broodiness in birds. The present study was aimed at identification of 24bp (insertion-deletion) indel polymorphism at the promoter region of prolactin gene and its association with broodiness in Telllicherry native chicken population. Methods: A total number of 200 birds of Tellicherry native chicken were randomly selected from All India Co-ordinated Research Project (AICRP) on poultry improvement, Mannuthy, Thrissur, Kerala. Blood samples were collected from the wing vein under aseptic condition and isolation of Genomic DNA was done. Isolated DNA samples were subjected to polymerase chain reaction (PCR) using specific set of forward and reverse primer to detect a 24bp indel polymorphism in the prolactin gene. PCR amplicons were subjected to further molecular analysis. Result: According to the polymorphic patterns birds were categorized to three different genotypes viz., II (insertion-insertion), ID (insertion-deletion) and DD (deletion-deletion). The genotype and allele frequency was calculated and the frequency of I allele (0.6975) was found to be higher than D allele (0.3025). Results of this study suggest that the incubation behaviour could be eliminated in Tellicherry native chicken population by increasing the frequency of I allele upon selective breeding which may enhance the egg production. Therefore, this 24bp indel polymorphism could be used as a molecular marker in poultry breeding.


2020 ◽  
Vol 17 (35) ◽  
pp. 174-180
Author(s):  
Salah H. FARAJ ◽  
Asaad Y. AYIED ◽  
D. K. SEGER

Several polymorphisms in different gene loci have been noted to affect production traits such as milk yield and milk composition. The present study aimed to determine the allelic and genotypic frequency of the DGAT1 gene and evaluate the associations between DGAT1 genetic variants and milk yield and its chemical composition of local Iraqi cattle. Blood samples from 100 cows were obtained for DNA isolation. The primer used in this study amplified 411-bp fragments at exon 8 of the DGAT1 gene. DNA sequencing methods were applied to detect single nucleotide polymorphism of the DGAT1 gene in 100 cows. The nucleotide sequences of exon 8 of the DGAT1 gene were registered for local Iraqi cattle in the National Center for Biotechnology Information (NCBI), DNA Data Bank of Japan (DDBJ), and the European Nucleotide Archive (ENA) under the following accession numbers (LC492073 and LC492074). The results showed the presence of two polymorphic sites leading to the construction of 2 different haplotypes in the cow. Haplotype diversity was 0.536, while nucleotide diversity was 0.0031. Two single-nucleotide polymorphism (SNP) loci of the DGAT1 gene were detected, namely A10433G (A/G) and A10434C (A/C). The resulting of this mutation changes lysine to alanine substitution at position 232 (A232K mutation) of amino acid sequence. Geneious software V. 2020.0.4 was used to detect genotypes of the DGAT1 gene, as the sequence alignment showed the presence of three genotypes. The genotypic frequencies of KK, KA, and AA were 0.40, 0.30, and 0.30, respectively. Frequencies of K and A alleles were 0.60 and 0.40, respectively. The KK genotype was significantly (P 0.05) associated with higher fat yield. Therefore, the DGAT1 gene could serve as a genetic marker for the selection of fat yield in cows.


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