拟穴青蟹凝集素基因Splec1的克隆及原核表达载体的构建

Cloning of Splec1 Gene from Scyiia paramamosain and Construction of Its Prokaryotic Expression Vector

为获得高表达拟穴青蟹(Scyiia paramamosain)凝集素(Lectin)Splec1的工程菌,本实验根据GenBank中凝集素的基因序列,设计合成1对扩增Splec1基因完整ORF的特异性引物,采用RT-PCR技术从拟穴青蟹血液中分离扩增了Splec1的cDNA序列(495 bp),将该基因重组到原核表达载体pET32a(+)中,酶切和测序分析表明,重组质粒pET32a(+)-Splec1结构正确.

Specific primers were designed according to the reported cDNA sequence of the Splecl, and the cDNA sequence of Splec 1 was obtained. Splecl sequence was then cloned to prokaryotic expression vector pET32a（ ＋ ）. The expression vector was checked then with two enzymes cutting and sequencing. Then recombination vector was constructed successfully.