苏云金杆菌cry IVD基因的克隆及序列分析

CLONING AND SEQUENCE ANALYZING OF B.t.i cryIVD GENE

目的 克隆苏云金杆菌 ( B.t.i) cry IVD基因并测定其序列。 方法 根据 B.t.i cry IVD已知序列 ,设计合成 1对引物 ,应用 PCR技术扩增 cry IVD基因 ,克隆入 p UC18载体 ,阳性克隆的重组质粒经酶切、电泳鉴定后进行序列测定。 结果 PCR扩增得到特异的 2 .0 kb基因片段。重组质粒经酶切后得到预期大小的基因片段。序列分析证明目的基因被完整且正向克隆。 结论 B.t.i cry IVD基因被成功克隆。

Objective To clone the Bacillus thuringiensis subsp.israelensi(B.t.i) cryIVD gene and determine its necleotide sequence. Methods A pair of primer was designed according to the known sequence of B.t.i. cryIVD. The B.t.i cryIVD gene was amplified by PCR technique and cloned into pUC18 vector. The positive clones were screened and identified by agarose gel electrophoresis and endonuclease digestion, then to determine its necleotide sequence. Results Identified by endonuclease digestion and sequence analysis, the B.t.i cryIVD gene fragment (2.1 kb) was specifically amplified. The recombinant plasmid was correctly constructed. Conclusion The B.t.i cryIVD gene was successfully cloned and sequenced.