蛋鸡OC-116基因的克隆和序列分析

参考原鸡（G. gallus） OC-116编码基因序列（登录号：NM_204569.1）设计3对引物，特异性地从仙居鸡子宫组织中克隆OC-116编码基因的N端、中段和C端，测序鉴定后通过酶切连接获得家鸡OC-116的全长编码基因。根据本研究的测序结果，初步发现突变概率较高的碱基有：第1233位（A→G）、1336位（C→G）、1358位（T→G）、1391位（T→G）、1491位（T→G）、1754位（G→T）、1841位（T→C）、1843位（C→T）、1983位（C→T）和2057位（T→C）；其中第1358,1754,1841,1983和2057位的碱基突变均为同义突变，第1233、1336、1391、1491和1843位是错义突变。而且突变碱基主要属于T：G颠换类型，其次是T：C转换类型。另外根据同源性分析，家鸡的OC-116编码基因在进化过程中很保守。总之，本研究已经从仙居鸡子宫组织中克隆到OC-116编码基因，为进一步研究该基因的功能奠定基础。

Cloning and Sequence Analysis of OC-116 gene of layers

According to the coding sequence of OC-116 （Ovocleidin-116） gene in the jungle fowl （GenBank No.： NM_204569.1） , we designed three pairs of primers to specially clone the N-terminal, the middle, and the C-terminal regions of OC-116 from the uterine cDNA of xianju layers. The sequencing results showed that we have achieved the full length coding gene of domestic chicken OC-116 by means of digestion and ligation. With the nucleotide sequence of jungle fowl OC-116 as reference, there were several mutations occurred in domestic chicken OC-116, among which, the mutated bases with high testing rate showed as follows,1233A→G,1336C→G,1358T→G, 1391T→G,1491T→G,1754G→T,1841T→C,1843C→T,1983C→T and 2057T→C.Among the above mutated bases,the 1358nt,1754nt,1841nt,1983nt,and 2057nt were synonymous mutations,and the others were missense mutations. Furthermore,most of the base mutations were belonged to the mutation forms of T ：G transversion, and T ：C transition.The gene homologous analysis show that chicken OC-116 gene is of highly evolutionary conservation. In a word,the present study has cloned the OC-116 coding gene from xianju layers which would facilitate the further investigation on the functional mechanisms of C-116 gene.