摘要
目的 构建和表达旋毛虫重组质粒。方法 PCR法在旋毛虫抗原基因Ts87两端加上合适的酶切位点和KOZAK序列,与真核表达载体pcDNA3.1(+)连接,构建pcDNA3.1/Ts87重组质粒。采用Lipofectamine介导的细胞外源DNA转染法,把重组质粒导入COS7细胞。分别用肌肉注射和基因枪免疫小鼠。结果和结论 成功构建pcD-NA3.1/Ts87重组质粒,并在COS7细胞和BALB/c小鼠体内表达。
Objective To construct and express DNA plasmid of Trichinella spiralis. Methods The BamHI and Hindlll enzyme sites and KOZAK sequences were introduced at both ends of Ts87 gene by PCR. Ts87 gene was ligated into pcDNAS. 1( + ) vector with T4 ligase. The recombinant plasmid pcDNAS. 1/Ts87 and plasmid pcDNAS. 1 were transfected into an eukaryotic cell line GOS7 through Lipofectamine, respectively. The BALB/c mice were immunized with the purified plasmid DNA pcDNAS. 1/Ts87 through two routes: intramuscular injection and gene-gun injection. Results and Conclusion The pcDNAS. 1/Ts87 was expressed both in COS7 and in the BALB/c mice.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2003年第2期93-95,共3页
Chinese Journal of Parasitology and Parasitic Diseases
基金
美国中华医学会(CMB)(Parasitology 98-674)
国家教委留学回国人员科研基金(1998~2001)
北京市教委科技发展基金(01KJ-091)
北京市跨世纪优秀人材工程(2001~2003)
河北省自然科学基金(302143)
