弓形虫主要抗原基因（SAG1）在昆虫细胞中表达的研究

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。

Study on the Expression of Toxoplasma gondii SAG1 Gene in Incect Cells

The gene encoding the major surface membrane protein (SAG1) was amplified and inserted into a Tn7 donor plasmid pFASTBAC1.The resulting recombinant FT9 was trans formed with Escherichia Coli DH10Bac competent cells containing baculovirus shuttle vectors (bacmids).By Tn7 transposition,SAG1 gene was integrated into the bacmid to form recombinant baculovirus.DNAs prepared from two such recombinants were trans fected with Spodptera frugiperda (Sf9) cells.A clear band correspondent to SAG1 was seen on the nitrocellulose membrane when immunoblotting using the rabbit antiserum against Toxoplasma gondii was performed to identify the trans-fected Sf9 cells.Thus,we believe that SAG1 protein can be expressed in the insect cells.