蛋鸡OC-116基因的克隆和序列分析

Cloning and Sequence Analysis of OC-116 gene of layers

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。

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.