摘要
用限制性内切酶ＰＳｔⅠ部分消解含有美洲鲽抗冻蛋白基因的ｐＣＴ５质粒，分离得到３２４ｂｐ的片段，将此片段亚克隆到ｐＵＣ１９质粒中，筛选到ｐＵＣ－ＡＦ重组子。从ｐＵＣ－ＡＦ重组子中，用ＢａｍＨⅠ和ＨｉｒｄⅢ切下３２４ｈｐ的抗冻蛋白基因，再重组到含有ＳＶ４０病毒启动子的ＰＫＳＶ－１０的载体中，构建成转基因鱼的表达载体，此表达载体可用于鱼的基因转移的研究。
Partially digesting plasmids pCT5 which contain antifreeze protein genes of Pseudopleuronects with restriction enzyme Pst I,the fragments of 324 base pairs are isolated and subcloned into plasmids PUC-19.Then recombinant plasmid pUC-AF is screened.After this,antifreeze protein genes of 324 base pairs are digested from plasmids pUC-AF with restriction enzyme BamH Ⅰ and Hind Ⅲ,and are inserted into plasmlds pKSV-10 which SV40 promoter.Finally,an expression vector is constructed for transgenic fish,and it may use in research of fish's gene transfer.
出处
《生物技术》
CAS
CSCD
1995年第4期34-36,41,共4页
Biotechnology
基金
黑龙江省科委"八五"项目
