人MASP-2N端片段的原核表达

Prokaryotic expression of the N-terminal Fragment of human mannose- binding lectin associated Serine Protein Kinase-2

目的克隆并表达人MASP-2N端片段，为制备单克隆抗体及其临床应用奠定基础。方法采用RT—PCR技术从人胎肝组织总RNA中扩增人MASP-2N端cDNA，克隆入pGEX-6p-2表达载体，在E．coli中表达GST—MASP-2N融合蛋白。结果酶切图谱分析和DNA测序分析表明，成功构建了含人MASP-2N端片段cDNA的重组质粒，并在大肠杆菌中表达了人MASP-2N端多肽片段。结论成功克隆表达了人MASP-2N端片段。

Objective To clone and express the human MASP-2N-terminal fragment in E. coll. Methotis Total RNA was extracted from human fetal liver tissue and MASP-2N cDNA was amplified by RT-PCR. The PCR product was cloned into pGEX-6p-2 vector. E. coli was transformed by the recombinant plasmid and GST- MASP-2N fusion protein was expressed. Results The constructed recombinant plasmids were identified by enzyme digestion and DNA sequencing. The recombinant protein was expressed in E. coll. Conclusion The N-terminal fragment of human MASP-2 was cloned and expressed.