家蚕龙角突变基因RAPD分子标记筛选及其克隆

Screening and Cloning of RAPD Marker of Longjiao Gene (K) in Silkworm

以家蚕龙角 (K)近等基因系第 1 1代龙角个体 (K/ +)和同蛾区正常个体 ( +K/ +K)为材料 ,用 1 77个随机引物进行PCR扩增 ,筛选出长度为 70 0bp的与龙角基因有关的特异性片段 (OPV 1 0 70 0 ) ,用另外 2个带龙角蚕品种和 2个非龙角蚕品种进行PCR扩增验证 ,进一步确认OPV 1 0 70 0 是与K基因有关的RAPD分子标记。回收纯化所获得的分子标记片段 ,以 pBluescriptⅡSK 质粒作载体 ,用T A互补法克隆 ,然后对重组子进行PCR和EcoRⅠ +HindⅢ双酶切鉴定 。

We screened the RAPD molecular marker (OPV 10 700 ) by comparing the PCR products amplified from two breeds (K/+ and + K/+ K) which are Longjiao (K) near isogenic lines(NILs).Using the other two longjiao breeds and two non Longjiao breeds as template for PCR amplification,the OPV 10 700 was confirmed as a Longjiao linkage marker.The marker furthermore was cloned into pBluescript Ⅱ SK vecter by the method of T A match.