恶性疟原虫裂殖子表面蛋白—2基因克隆及其在大肠杆菌中 …

目的：克隆恶性疟原虫裂殖子表面蛋白２全基因，并在大肠杆菌中进行表达。方法：采用ＰＣＲ扩增恶性疟原虫ＦＣＣ－１／ＨＮ株ＭＳＰ２基因，克隆插入ｐＵＣ１９，并重组人表达质粒载体中，转化大肠杆菌，诱导重组质粒ｐＢＫ－ＣＭＶ／ｍｓｐ２表达ＭＳＰ２蛋白抗原，对表达产物进行ＳＤＳ－ＰＡＧＥ，ｄｏｔ－ＥＬＩＳＡ和Ｗｅｗｔｅｒｎ ｂｌｏｔ鉴定。结果：ＰＣＲ扩增出８２４ｂｐＤＮＡ片段，经酶切鉴定和部分序列测定证实为Ｍ

Cloning of merozoite surface protein-2 gene of Plasmodium falciparum and its expression in E. coli

Objective: To clone whole merozoite surface protein-2(MSP2) gene of Plasmodium falciparum and express it in E. coli. Methods: Whole MSP2 gene fragment was amplified from the genome DNA of Plasmodium falciParum FCC-1/ HN strain by PCR and was inserted into plasmid pUC19 and then subcloned into the expressing plasmid. The recombinant vectors were transformed into E. coli. The transformed bacteria bearing pBK-CMV/msp2 plasmids were induced for the production of MSP2. The expressed product was analyzed with SDS-PAGE, dot-ELISA and Western blot. Results: A fragment sized 824 bp was amplified by PCR. It was confirmed by restriction site mapping and sequencing that the fragment was MSP2 gene. pBK-CMV/msp2 transformed bacteria produced the desired protein with a relative molecular mass of 3. 2 * 10~4, presenting 10% of the total bacterial proteins. The expressed protein exhibited a strong reaction with anti-malarial antibodies as detected by dot-ELISA with serum obtained from patient with falciparum malaria. When the products were blotted with the same serum, specific bands of 3. 2 *10~4 representing MSP2 were identified. Conclusion: MSP2 gene of Plasmodium falciparum can be expressed in E. coli. But more efficient expression system for MSP2 should be established.