重组质粒PcDNA3.1/sTNFRⅠ在小鼠体内表达的初步研究

A preliminary study on the expression of recombinant plasmid PcDNA3.1/sTNFRⅠ in mice

目的检测构建的PcDNA3.1/sTNFRⅠ真核表达载体在小鼠体内能否有效表达目的产物,为探索利用该载体基因治疗TNFα介导的炎性疾病奠定一定的实验基础。方法将PcDNA3.1/sTNFRⅠ载体直接注入小鼠胫前肌,ELISA法检测肌肉匀浆中的sTNFRⅠ表达。结果在注射后7d与10d,PcDNA3.1/sTNFRⅠ注射组小鼠肌肉匀浆中sTNFRⅠ表达的量均显著高于PcDNA3.1注射组(P<0.01),注射后第10天sTNFRⅠ表达量高于第7天sTNFRⅠ表达量(P<0.01)。结论采用直接注射法将PcDNA3.1/sTNFRⅠ载体转移入肌肉内可以使目的基因得到一定的表达。

Objective： To explore whether the recombinant plasmid PcDNA3. 1/ sTNFR Ⅰcould be expressed properly in mice. Methods：The recombinant plasmid PcDNA3.1/sTNFR Ⅰ and the empty vector PcDNA3.1 were directly injected into the fight tibialis anterior muscle of each 6 mice respectively. The expression product, sTNFRⅠ , in muscle homogenates was measured by ELISA 7 and 10 days after injection respectively. Results：7 days after injection sTNFR Ⅰ （mg/g） in muscle homogenates in PcDNA3.1/sTNFR Ⅰ and PcDNA3.1 injected mice was 161. 123 ±4.0 and 69.34 ±1. 489（ P 〈0.01 ）, 10 days after injection 191. 727 ± 3. 361 and 77. 664 ± 2. 927（ P 〈0.01 ） ,respectively. In each group 10 days after injection sTNFR Ⅰ level was higher than 7 days after injection （P 〈0.01 ）. Conclusion：Recombinant plasmid PcDNA3.1/sTNFR Ⅰ can be expressed in muscle of mice.