细菌感染后大黄鱼脾脏组织中PcQM基因表达及PcQM在大肠杆菌中原核表达

本研究利用Real-time PCR方法,分析利用哈维氏弧菌和副溶血弧菌混合菌液攻毒12 h、24 h和48 h后,大黄鱼脾脏组织中PcQM基因表达的变化情况。结果表明,攻毒12 h后脾脏组织中大黄鱼PcQM基因表达量显著降低0.031±0.006,且差异极显著（p〈0.01）;攻毒24 h后,表达量显著增加16.240±2.822倍,且差异显著（p〈0.05）;攻毒48 h后脾脏组织中大黄鱼PcQM基因表达量显著降低0.115±0.025（p〈0.01）。初步推测,PcQM基因在大黄鱼免疫系统中起着重要的调控作用。本研究构建了PET28a（＋）/PcQM表达载体,进行体外表达。用Western-blotting分析,证实有一条约为25 kD的蛋白条带,表明PcQM蛋白体外表达成功。本研究为进一步研究PcQM蛋白免疫调节功能奠定了基础。

The Expression Profiles of the Pc QM Gene in the Spleen Tissue of Infected Pseudosciaena crocea and Prokaryotic Expression of the PcQM in E. coli

In this study, the expression profiles of the PcQM gene was analyzed by Real-Time PCR method in the spleen tissue ofPseudosciaena crocea that was infected by bacteria of Vibrio harveyi and V. parahaernolyticus after 12 h, 24 h and 48 h. The results showed that expression level of the PcQM gene was obviously down-regulated at 12 h post-challenged, and the expression was 0.031±0.006 （P〈0.01）; it was increased at 24 h post-challenge, and the expression was 16.240＋2.822 （p〈0.01）; it was significantly down-regulated at 48 h post-challenge, and the expression was 0.115±0.025 （p 〈0.01）, which suggested that the PcQM gene should have an immunomodulatory effect on P. crocea. PET-28a （＋）/PcQM was constructed and expressed in vitro, and a 25 kD recombinant fusion protein with a His tag was confirmed by the Western-blotting, which showed that prokaryotic expression vector of PcQM was constructed and expressed successfully. The study laid a foundation for further investigation of the im- mune regulation function of PcQM protein.