新孢子虫与弓形虫交叉抗原基因AMA1的克隆及原核表达载体构建

Clone and prokaryotic expression vector construction of Neospora caninum and Toxoplasma gondii AMA1 gene

为了解新孢子虫与弓形虫交叉抗原基因AMA1的生物学特性,应用PCR技术扩增新孢子虫/弓形虫AMA1基因,将PCR产物纯化回收后克隆于pMD18-T Simple Vector,构建pMD18-T-Nc/Tg AMA1重组克隆质粒,经PCR鉴定、酶切鉴定及测序分析后,亚克隆于原核表达载体pGEX-4T-1,构建重组原核表达质粒pGEX-4T-Nc/Tg AMA1,经PCR鉴定、酶切鉴定表明,构建的原核表达载体正确。该试验为新孢子虫/弓形虫AMA1基因重组疫苗的研究奠定了基础。

In order to understand the biological characteristics of Neospora and Toxoplasma＇ s antigen gene AMAI,we was amplified Neospora/Toxoplasma AMA1 gene by PCR, purified the product of PCR and connected to the cloning vector pMD18-T simple vector to con- struct pMD18 -T-Nc/Tg AMA1 recombinant plasmid clones. The expression plasmid which is identifieated correctly by PCR and enzyme digestion and sequencing analysis will be connected to prokaryotic expression vector pGEX-4T-1, eventually build prokaryotic expression vector pGEX-4T-Nc/Tg AMA1. It will provide a foundation for the study of Neospora/Toxoplasma AMA 1 recombinant vaccine.