摘要： 首先提取苦瓜的基因组DNA,然后根据Genbank中已公开发表的MAP30序列,设计一对特异引物MAP301,MAP302,并增添特异定位于内质网的小肽Kozak序列。采用PCR技术,从苦瓜的基因组DNA中扩增出一分子量为800bp左右的片段,回收克隆测序,结果表明,克隆获得片段与已公开发表的MAP30序列同源性达100%,将阳性克隆进行酶切,获得的目标片段与用同样酶切所获得的植物表达载体pEV1和pEV2大片段连接,酶切鉴定重组子构建成特异表达于果实的载体。

关键词: 苦瓜, MAP30, 载体构建, PCR

Abstract: Apair of primers(MAP301/MAP302)was designed based on the reported sequence of MAP30 in GenBank,and was then used to amplify the genomic DNA extracted from bitter melon by PCR,from which a 800 bp fragment was obtained.The fragment was cloned and then transformed to the E.coli DH5α after ligating with pMD18-T vector.The sequencing result of cloned fragment showed that it had 100% base similarity to the MAP30 gene.The cloned fragment was ligated with pEV1 or pEV2 vectors,and the expression vectors were construted after screening restriction analysis.The result provides the foundation for the further research on tomato as plant oral vaccine.

Key words: Bitter melon, MAP30, Construction of vector, PCR

中图分类号: 

• S642.50.6