转座子K1.4可提高荧光素酶基因在转基因家蚕中的整合频率  

Transposon K1.4 Can Increase the Integration Frequency of Luciferase Gene in the Transgenic Silkworm, Bombyx mori

作　　者：钱信果[1] 周丛照[1] 李振刚[1] 

机构地区：[1]中国科学技术大学生物系

出　　处：《中国科学技术大学学报》1998年第6期710-713,共4页JUSTC

基　　金：安徽省科委"九五"攻关项目

摘　　要：合适的整合表达载体对于获得转基因家蚕具有重要意义，为此本文构建了包含家蚕转座子Ｋ１．４、丝素基因上游启动子序列和荧光素酶基因ｃＤＮＡ的质粒ＳＫＦＬ以及不含Ｋ１．４的质粒ｐＦＬ，并将其通过扎卵法转移到家蚕受精卵中．通过对当代转基因家蚕５龄幼虫丝腺和蚕体ＤＮＡ进行斑点杂交和Ｓｏｕｔｈｅｒｎ杂交发现，荧光素酶基因在家蚕不同组织中的整合是随机的，但携带Ｋ１．４的质粒ＳＫＦＬ的整合频率明显高于质粒ｐＦＬ．这说明转座子Ｋ１．４可以有效提高外源基因在转基因家蚕中的整合频率，从而有可能成为构建转基因家蚕的一种有用的整合载体．In order to study the effect of transposon K1.4 on foreign genes' integration frequency, a recombinant plasmid SKFL was constructed which contained Fb(the 5'-flanking region from -850bp to +75 of the fibroin heavy chain of Bombyx mori), transposon K1.4 and luciferase gene (Luc), used as a report gene. The other recombinant plasmid pFL containing Fb and Luc was constructed as control. The fresh Bombyx mori eggs laid in 3-8 hous were divided into two groups: one was transferred with SKFL as an anexperimental group and the other was transferred with pFL as control group. After the surviving eggs were raised to the fifth instar larvae, DNAs, RNAs from the silkglands and the bodies of silkworms were extracted respectively. DNA Dot Blot and Southern Blot indicated that luciferase gene had been integrated into the genome of Bombyx mori in the experimental group, but the integration sites of luciferase gene distributed randomly, which accorded with the mosaicism of the foreign genes* integration in host cells. However, luciferase gene did not integrate in the genome of Bombyx mori in the control group. These results provided us with the evidence that K1.4 played an important role in increasing the integration frequency of the foreign gene. Moreover, the negative result of RNA Dot Blot indiated that luciferase gene did not express in the fifth instar larvae of Bombyx mori. Nevertheless, the transposon K1.4 could be a very useful integration vector in constructing transgenic silkworms.

关 键 词：家蚕 转座子 荧光素酶基因 整合频率 转基因家蚕 

分 类 号：S882[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]