title: [Applications of SSCP and HMA for polymorphic analysis of horse MHC-I alleles].
abstract: In this article, we report the analysis of genetic polymorphisms of horse MHC-I molecules by SSCP and HMA, which are methods based on the technique of polyacrylamide gel electrophoresis (PAGE). Our results showed that SSCP was not a suitable method for the analysis of genetic polymorphisms of horse MHC-I molecules due to the failure in generating satisfied separation of DNA fragments, even if experimental conditions were optimized. However, the HMA method produced clearly separated DNA fragments of horse MHC-I molecules, after the experimental conditions, such as the running temperature and the concentration of detergent, were optimized by using a reference plasmid. PCR-amplified samples from horses No. 6, No. 7, No. 8, No. 9 and No. 10 generated 6, 5, 6, 5, and 7 bands, respectively, in corresponding lanes of the polyacrylamide gel. DNA fragments in each band cut from the gel were amplified by PCR using a second pair of primers, and were cloned for sequencing. Alignment analysis of these sequences revealed that HMA was a proper method to efficiently analyze the polymorphisms of MHC-I molecule genes.
authors.affiliation: Institute of Animal Husbandry, Shenyang Agricultural University, Shenyang 100161, China. xiangwei1981213@163.com
authors.forename: Wei
authors.initials: W
authors.lastname: Xiang
authors.forename: Jian
authors.initials: J
authors.lastname: Ma
authors.forename: Xue-Feng
authors.initials: XF
authors.lastname: Wang
authors.forename: Yu-Jun
authors.initials: YJ
authors.lastname: Zhao
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Zhou
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006736:Horses / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational / Q000235:genetics*
chemical_list: D015395:Histocompatibility Antigens Class I