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用PCR技术从弓形虫DNA扩增出一段长约902bp编码弓形央主要膜蛋白抗原(SAG1)的基因,将SAG1基因插入带有转座子供体的pFastBacl质粒,得到重组质粒pFT9,pFT9转化含有杆状病毒穿梭载体的DH10Bac感受态细胞,通过转座重组作用使SAG1基因整合到杆状病毒载体上,将两株转座重组子BaC101和Bac304DNA分别转染Sf9昆虫细胞,用弓形虫免疫血清所作的免疫印迹试验证实这两株重组杆状病毒的昆虫细胞株均表达了SAG1蛋白。本文首次报告弓形虫SAG1蛋白在昆虫细胞中的表达。
PCR was used to amplify a DNA fragment of approximately 902 bp encoding the major archival protein A (SAG1) from the Toxoplasma gondii DNA. The SAG1 gene was inserted into the pFastBac1 plasmid with the transposon donor to obtain the recombinant plasmid pFT9, Like virus shuttle vector DH10Bac competent cells, the SAG1 gene was integrated into the baculovirus vector by transposon recombination, the two transposon recombinant BaC101 and Bac304 DNA were transfected Sf9 insect cells, with Toxoplasma gondii immune serum made Western blotting confirmed that SAG1 protein was expressed in the insect cell lines of these two recombinant baculoviruses. This is the first report of Toxoplasma gondii SAG1 protein expression in insect cells.