摘要
目的 克隆日本血吸虫 31/ 32 KDa蛋白质的编码基因 ,用于血吸虫病免疫诊断和预防。方法 以日本血吸虫成虫 RNA为模板逆转录合成 c DNA链 ,参照 S.m 31/ 32编码序列设计合成引物 ,用 PCR法扩增 31/ 32 KDa蛋白质基因编码序列 ,将其克隆入 p GEM- T载体 ,并用双酶切和以质粒为模板的 PCR进行鉴定。结果 RT- PCR扩增出一条约 12 70 bp大小的特异性条带 ,重组质粒的双酶切和以质粒为模板的 PCR均获得了一条与 RT- PCR扩增出的条带大小相同。结论 日本血吸虫 31/ 32 KDa蛋白质重组 p GEM- T克隆载体的成功构建 ,为进一步研究提供条件。
Objecitve To clone 31/32 KDa proteins gene of Schistosoma japoninicum in order to study its effect on immunological diagnosis and prevention. Methods Synthesis of the first SjcDNA strand was driven by superscript Ⅱ RT using adult worm RNA as template. Specific primers was designed and synthesized according to mRNA sequence of S.m 31/32KDa gene. Coding region gene of Sj 31/32 KDa proteins was amplified by PCR. The product from PCR was cloned into pGEM T 31/32 vector which was identified later by restriction analysis and PCR. Results For RT PCR, a specific band about 1 270 bp was amplified. The same band was obtained by double restriction of recombinant plasmids and PCR of using recombinant plasmids as template. Conclusions pGEM T 31/32 KDa was successfully constructed and provided as the basis for further studying on 31/32 KDa proteins expression and its function.
出处
《疾病控制杂志》
2000年第4期302-306,共5页
Chinese Journal of Disease Control and Prevention
基金
安徽省科委科研基金!资助 (0 0 2 2 0 31)
安徽省教委科研基金!资助 (98JL0 6 2 )
