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Study on Polymorphysm of MC4R Gene and Its Relationship with Early Growth Traits of Hu Sheep

Author: ZhangGaoZhen
Tutor: JiangYongQing；ShiFangXiong
School: Nanjing Agricultural College
Course: Animal Genetic Breeding and Reproduction
Keywords: Hu sheep growth curve MC4R gene PCR-SSCP
CLC: S826
Type: Master's thesis
Year: 2009
Downloads: 22
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Abstract


In the current study, early growth and development pattern of Hu sheep was studied. In addition, Melanocortin receptor-4(MC4R) gene which related to growth traits was selected as a candidate gene. By Bioinformatics method, the MC4R gene of Hu sheep was cloned to get the DNA sequence. By PCR-SSCP analysis, the relationship between single nucleotide polymorphism and early growth traits in Hu sheep were used to screen out the molecular marker associated with growth traits.1. The Fitting of Early Growth Curve of Hu SheepVon Bertalanffy model was best fitting in the early growth curve of Hu sheep among the three nonleaner models (Logistic, Von bertalanffy and Gompertz). According to the Von Bertalanffy model, the growth curve in early growth period of Hu sheep was fitted. It also provided the theoretical reference base for further breeding and improving management of feeding.2. The Cloning and Bioinformatics Analysis of MC4R Gene of Hu sheepMC4R gene of Hu sheep was cloned. Its DNA was 1765bp in length, including of 5’-UTR(213bp),3’-UTR of 553bp and ORF(999bp), which codes 332 amino acids. The molecular weight of MC4R proteins in Hu sheep was 36.57 kDa and its isoelectric point was 7.09. MC4R protein of Hu sheep also contained the transmembrane regions. The MC4R protein sequence of Hu sheep had 95.80%,91.87%,91.57%,90.96% and 90.36% idential to the cow, dog, human and rhesus, rat and mouse, individually. The putative MC4R protein of Hu sheep contained many fuctional site and motif. The result revealed that this protein had conserved structure with other MC4R.3. Detection of MC4R gene Mutation with PCR-SSCP and the Association between Its Polymorphisms and Growth CharacterThe mutation and DNA sequence of MC4R gene in Hu sheep was determined in the current study using PCR-SSCP. Two SNPs were detected in amplification sequence of primer M6 and M7 among eight primers, and each of them behaves 3 genotypes. Allele B majors in SNPM6.Allele frequencies of allele B and A were 0.7361 and 0.2639, respectively; Allele C majors in SNPM7.Allele frequencies of allele C and D were 0.7292 and 0.2708, respectively. Theχ2 fitness test suggested that the population were in the state of Hardy-Weinberg equilibrium at SNPM6 and in state of disequilibrium at SNPM7.The PIC (Polymorphism Information content) of SNPM6 and SNPM7 were 0.313 and 0.3169, respectively, ranked at middle levels of polymorphism. This result revealed that SNPM6 and SNPM7 had more genetic variation and expected to have more choices.Mean and standard error of least squares results showed that the weaning weight of genotype AA of SNPM6 was extremely significantly higher than that of BB and AB (p<0.01), the weaning weight of genotype DD of SNPM7 was extremely higher than that of CC and CD (p<0.01), the birth weight of genotype DD of SNPM7 was extremely higher than that of CC (p<0.01) and significantly higher than that of CD (p<0.05).Allele A of SNPM6 and allele D of SNPM7 have positive effect on weaning and birth weight of Hu sheep, however, Allele A of SNPM6 and allele D of SNPM7 had negative effect on weaning and birth weight of Hu sheep. Result of correlation Analysis showed that the highly significant correlationship between SNPM6 and weaning weight of Hu sheep was observed. SNPM7 both showed a highly significant correlation with weaning weight of Hu sheep.In conclusions, the results of our current project suggest that the MC4R gene be a candidate gene close correlated to early growth traits in Hu sheep. The SNPs detected in MC4R gene of Hu sheep is a good marker assisted selection programs for growth traits in Hu sheep to accelerate breeding progress.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Sheep
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