弓形虫主要表面抗原基因编码序列的扩增、克隆及原核融合表达

AMPLIFICATION, CLONING AND PROKARYOTIC EXPRESSION OF GENE FOR MAJOR SURFACE ANTIGEN OF TOXOPLASMA GONDII

目的扩增弓形虫主要表面抗原（P30）基因编码序列并进行重组表达方法设计合成引物，从弓形虫基因组DNA中扩增P30基因编码序列，克隆入载体pGEM-T和PGEX-4T-1，经PCR和酶切筛选，测序验证后，进行诱导表达和Westemblot鉴定结果从弓形虫基因组DNA中扩增出P30基因编码序列，并诱导表达出能被兔抗弓形虫血清识别的重组P30结论P30克隆载体和GST融合表达载体的构建和P30的成功表达，为进一步从基因水平和蛋白水平研究P30及进行诊断试剂和疫苗研究创造了条件。

Aim To amplify P30 gene and express P30 fusion wiht GST Methods P30 gene was smplified from T, gondii chromosomal DNA and liated to pGEM-T and pGEM-4T-1. Screening-positive recombinants were induced for expres-sion, which was subsequently WB Results P30 gene was amplified and GST-fusion was confirmed by rabbit antiT. gondii serum.Conclusions The construction of pGEM-T-P30 and pGEX-4T-1-P30, together with the recombinan protein would lay a base for further investigation of P30 at a mole-level and application to diagnosis and vaccination