Study On Genetic Variation And Its Effects And Expression Of CFL2 Gene In Chickens

CFL2 gene is a member of the cofilin family of actin-binding proteins,which affects muscle development and myofibrillar formation.In this study, the genetic variations of CFL2 gene were detected by DNA pool sequencing. SNPs of CFL2 gene were genotyped in an F2 resource population of Gushi chickens crossing with Anka broilers using PCR-RFLP and CRS-PCR-RFLP. The associations of 2 SNPs polymorphisms with some parameter in 849 F2 chickens were analyzed. The partial coding sequence of Silky Chicken's CFL2 gene was cloned and analyzed by bioinformatics methods. Using real-time PCR techique, the mRNA of CFL2 gene in different tissue (heart, liver and leg muscle ) and different week (0,4,8and 12 week)were analyzed. The goal of this study is to identify the sequence variations of the CFL2 gene to analyze the relationship between gene variation and chicken performance, and to find molecular markers for chicken genetics and breeding programs. The main results were summarized as below:1. Three SNPS were found in CFL2 gene with the location of intron 1 (T853G) and 3'UTR(T2484C,G2545A), respectively. T2484C and G2545A were newly discovered. T853G polymorphisms was significantly associated with semi-evisceration weight, breast muscle fiber density,γglutamyltranspeptidase, alkaline phosphatase and triglyceride(P<0.05); G2545A polymorphisms was significantly associated with shank girth and shank length at 4 weeks,and with leg muscle fiber density and leg muscle fiber diameter.2. The partial coding region sequence of CFL2 gene in Silky fowl were obtained. It has 12 phosphorylation sites and a conserved domain of ADF but no signal peptide sequence. There are 4 hydrophobic regions . The amino acid sequence contains four kinds of secondary structure:α-helix,β-fold,β-angle and random coil but no transbilayer helix.It is a highly conserved protein.3. The highest expression of CFL2 was found in leg muscle and the lowest in liver. The expressions of CFL2 in leg muscle,heart and liver was significantly difference(P<0.05),and the expressions of CFL2 in leg muscle,heart and liver was highest in 8 weeks , respectively.