Pollack Y, Shemer R, Metzger S, Spira D T, Golenser J
Exp Parasitol. 1985 Dec;60(3):270-5. doi: 10.1016/0014-4894(85)90031-1.
Genomic libraries of Plasmodium falciparum were constructed in the pBR322 plasmid. Using the DNA-mediated gene transfer technique, the genomic libraries were introduced into tissue-cultured mouse cells lacking the enzyme adenine phosphoribosyltransferase. Following selection for the adenine phosphoribosyltransferse phenotype, several colonies were isolated. All clones were shown to possess adenine phosphoribosyltransferase activity and pBR322 sequences. In addition, the Km value of adenine phosphoribosyltransferase (for adenine) from a transformant was found to be identical to that from P. falciparum. These results indicate that the adenine phosphoribosyltransferase gene of P. falciparum was successfully cloned and expressed in a mammalian system.
恶性疟原虫的基因组文库构建于pBR322质粒中。利用DNA介导的基因转移技术,将基因组文库导入缺乏腺嘌呤磷酸核糖转移酶的组织培养小鼠细胞。在选择腺嘌呤磷酸核糖转移酶表型后,分离出了几个菌落。所有克隆均显示具有腺嘌呤磷酸核糖转移酶活性和pBR322序列。此外,发现来自转化体的腺嘌呤磷酸核糖转移酶(针对腺嘌呤)的Km值与恶性疟原虫的Km值相同。这些结果表明,恶性疟原虫的腺嘌呤磷酸核糖转移酶基因已成功克隆并在哺乳动物系统中表达。