摘要
用CTAB法提取豇豆叶片总DNA后,根据基因两端的保守序列设计引物,经PCR方法扩增得到CpTI基因,将其克隆到pMD-18T载体上,酶切并测序鉴定,将该基因登陆到GeneBank(NO.EU088405)并进行同源性鉴定,证明该基因为丝氨酸蛋白酶抑制剂基因家族中的一员。在此基础上,将CpTI基因用限制性内切酶BamHⅠ/SacⅠ切下后,克隆到pCUbi1303的UBI启动子和NOS终止子之间。经PCR和酶切鉴定,得到了该基因的真核表达载体pCUbiCpTI1303,为下一步的转基因做好了准备。
Genomic DNA in the leaves of the cowpea was extracted with CTAB.The CpTI gene was amplified by PCR with the conserved sequence acted as primers,then cloned to pMD-18T vector and detected the recombinant by restriction enzyme method.The sequence has been registered in GeneBank（NO.EU088405）.It is one of serine protease inhibitor gene family.After digesting by BamHⅠand SacⅠ,the gene was cloned to pCUbi1303 between the promoter UBI and the terminator NOS.After detected by restriction enzyme and PCR,it indicated that the plant expression vector pCUbiCpTI1303 was constructed successfully,which provides a basis for the following transgene.
出处
《西北农业学报》
CAS
CSCD
北大核心
2008年第3期16-19,共4页
Acta Agriculturae Boreali-occidentalis Sinica
基金
教育部“新世纪优秀人才”支持计划
