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Zhang T.,Yangzhou University | Zhang T.,Key Laboratory for Animal Genetics | Zhang G.X.,Yangzhou University | Zhang G.X.,Key Laboratory for Animal Genetics | And 13 more authors.
Genetics and Molecular Research | Year: 2015

The gonadotropin-releasing hormone (GnRH) plays an important role in the control of reproductive functions. Recent studies have reported the occurrence of GnRH molecular variants in numerous species. In this study, the GnRH1 gene from Jinghai yellow chicken was cloned by reverse transcriptase-polymerase chain reaction and transformed into BL21 (DE3) competent cells. The GnRH1 gene and amino acid sequences were subjected to bioinformatic analyses. The GnRH1 gene nucleotide sequence was discovered to be 352 bp long, containing a coding, promoter, and section of the 3'-regions. The GnRH1 gene shared 93, 81, 54, 58, 61, 76, 76, 59, 76, and 66% sequence identity with Meleagris gallopavo, Columba livia, Homo sapiens, Bos taurus, swines, Capra hircus, Ovis aries, Pantholops hodgsonii, Equus caballus, and Rattus norvegicus, respectively. The GnRH1 gene showed conserved domains. The GnRH1 protein was a secreted protein comprising 92 amino acids, with a molecular weight of 10205.6 Da and a theoretical pI of 5.67. Most of the amino acid residues were observed to be hydrophilic, indicating water solubility. The predicted secondary structures of proteins included α-helices (h; 23.08%), β-extensions (e; 10.92%), and random coils (c; 66.0%). The successful construction of prokaryotic expression vector pET32a-GnRH1 was confirmed by restriction and sequence analysis. SDS-PAGE analysis showed the successful expression of recombinant plasmid in Escherichia coli BL21 (molecular weight = 25-28 kDa). Larger quantities of protein were expressed in supernatant, indicating greater expression in soluble form. Western blot analysis confirmed the expression of the target protein. © FUNPEC-RP.

Zhang T.,Yangzhou University | Wang W.,Yangzhou University | Zhang G.,Yangzhou University | Wang J.,Yangzhou University | And 2 more authors.
Yi chuan = Hereditas / Zhongguo yi chuan xue hui bian ji | Year: 2015

Body weight traits are important economic characters of broilers. This study was carried out to screen for molecular markers and candidate genes that can be used to improve the body weight traits. A herd of 400 female Jinghai yellow chickens were measured for body weights from 0 to 14 weeks of age. Genome-wide association study (GWAS) was carried out using specific-locus amplified fragment sequencing (SLAF-seq) technology to detect SNPs associated with body weight traits of Jinghai yellow chicken. Finally, 100 SNPs that associated with body weight traits were detected. The results showed that effects of 15 SNPs reached 5% Bonferroni genome-wide significance and 85 SNPs reached potential genome-wide significance. Genes in the candidate regions with 1 Mb windows (SNP position±0.5 Mb) surrounding each significant SNP were screened. Finally, nine candidate genes were obtained, among which four genes of FAM124A (Family with sequence similarity 124A), QDPR (Quinoid dihydropteridine reductase), WDR1 (WD repeat domain 1) and SLC2A9 (Solute carrier family 2 (facilitated glucose transporter), member 9) might be important candidate genes influencing body weight traits of Jinghai yellow chicken. Furthermore, it was also found that most SNPs associated with mid and late growth and body weights were intensively located in the region of 75.6-80.7 Mb on chromosome 4. Our study thus provides a basis for genetic understanding of the Jinghai yellow chicken body weight traits.

Tang Y.,Yangzhou University | Tang Y.,Key Laboratory for Animal Genetics | Zhang T.,Yangzhou University | Zhang T.,Key Laboratory for Animal Genetics | And 13 more authors.
Molecular Biology Reports | Year: 2014

The objective of this study was to analyze possible associations between single nucleotide polymorphisms (SNPs) in the Myf5 gene with chicken growth and reproductive traits. SNPs in Myf5 of the Jinghai yellow chicken were detected by the polymerase chain reaction single-strand conformation polymorphism method and the haplotypes were analyzed. Eight SNPs were identified in the exons of Myf5. Nine haplotypes were established in a group of 379 Jinghai yellow chickens. In terms of growth traits, least square analysis showed that haplotype H1H5 had significant effects on weight at weeks 8 and 12 (P < 0.05). Haplotype H2H6 had significant effects on weight at weeks 12 and 14 (P < 0.05). For reproductive traits, H1H5 had higher body weight for the first egg than H1H4 and H2H4 (P < 0.05), and H1H3 (P < 0.01). H1H3 had a poor performance in average egg weight at 300 days. On the other hand, H1H3 had an advantage in egg number at 300 days. The results showed that SNPs of Myf5 have certain effects on growth and reproductive traits in Jinghai yellow chickens, which can be used in marker-assisted selection to accelerate chicken genetic progress. © 2014, Springer Science+Business Media Dordrecht.

Yang F.,Yangzhou University | Jin C.,Yangzhou University | Dai G.,Yangzhou University | Xie K.,Yangzhou University | And 2 more authors.
Journal of Animal and Veterinary Advances | Year: 2012

In this study, exon 2 of Insulin-like Growth Factor Receptor type I (IGF-IR) gene was studied as a candidate gene associated with production performance in Jinghai Yellow chicken. A pair of primers was designed to screen the Single Nucleotide Polymorphisms (SNPs) using DNA pooling which consisted of DNA from all 200 female chickens and three pairs of primers were then designed to check them individually. The result showed there were three SNPs (G26333A, G26336A and C26639T) in the chicken group, each had three genotypes and was in Hardy-Weinberg equilibrium (p>0.05). Correlation analysis showed the G to A transition at 26333 bp was negatively associated with 300 days egg number and body weight at 4 and 16 weeks of age, the G to A transition at 26336 bp was positively associated with body weight at 8 and 12 weeks of age and the C to T transition at 26639 bp was positively associated with body weight at 16 weeks of age. Com-genotype analysis showed that the age at the first egg of chickens with AA/CD/EE was earlier than those with AA/CC/EE and AB/CC/EF (p<0.05) and chickens with AB/CC/EE had more 300 days egg number than those with BB/CD/EE (p<0.05). AA/CC/EF was an enhancing com-genotype for body weight while AA/CC/EE and AB/CC/EF were the negative ones at 12 and 16 weeks of age. The results suggested a significant association between SNPs of IGF-IR gene and production traits and these SNPs could be used as potential genetic markers in future breeding of Jinghai yellow chicken. © Medwell Journals, 2012.

Zhao X.H.,Yangzhou University | Zhao X.H.,Molecular Design International, Inc. | Wang J.Y.,Yangzhou University | Wang J.Y.,Molecular Design International, Inc. | And 5 more authors.
African Journal of Biotechnology | Year: 2011

In this study, insulin-like growth factor binding protein 2 (IGFBP-2) and signal transducers activators of transcription 5b (STAT5b) gene were studied as candidate gene associated with body weight and reproductive traits of the Jinghai Yellow chicken. Single nucleotide polymorphisms (SNPs) of the IGFBP-2 and STAT5b genes were examined in both Jinghai Yellow chicken and three reference chicken populations by PCR-SSCP. Two SNPs (T3746TG and CC3753TT) were detected in the IGFBP-2 gene. One SNP (C8066T) was observed in the STAT5b gene. For primer 1, the general linear model analysis showed that Jinghai yellow chickens with FF genotypes had significant effect on hatch weight, egg weight at 300 days and body weight at 300 days than those of the EF genotype and had significant effect on body weight at 8 weeks of age than those of the EE genotype (P < 0.05). For primer 2, Jinghai yellow chickens with CT genotype had significant effect on hatch weight and age at first egg than CC genotype and TT genotype respectively (P < 0.05). SNPs in both IGFBP-2 and STAT5b genes had significant effect on body weight and reproductive traits of the Jinghai yellow chicken than those with either SNP alone. These SNPs may be served as a potential genetic marker for growth and reproduction traits evaluation of the Jinghai yellow chicken. © 2011 Academic Journals.

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