盐度胁迫下三疣梭子蟹Ferritin基因的原核表达

Prokaryotic expression of the Portunus trituberculatus Ferritin gene during salinity stress

为验证Ferritin基因是否对三疣梭子蟹的盐度胁迫具有适应性,利用PCR扩增获得了三疣梭子蟹(Portunustrituberculatus)第六对鳃的Ferritin基因,将该目的基因克隆到表达载体pET28a(+)中,转化大肠杆菌DE3(BL21),并进行IPTG诱导表达。结果表明:经诱导的转pET28-Ferritin重组质粒菌株有特异的表达蛋白条带出现,与预期的理论值(19.448 kD)相符合;高盐胁迫下,转重组质粒的大肠杆菌比转pET28空载质粒存活率显著提高(P<0.001),并且随盐度升高,两者差异越显著。这表明Ferritin基因能够提高大肠杆菌的盐度耐受力,由此推测Ferritin基因参与了三疣梭子蟹盐度适应的生理过程。本研究旨在为生产过程中三疣梭子蟹养殖水体的盐度调控策略提供理论依据。

We evaluated the effect of ferritin on the ability of the swimming crab to adapt to salinity stress. We amplified the Ferritin gene from the sixth gill of the swimming crab and cloned it into the pET 28 a （＋） expression vector. The vector was transformed into Escherichia coli DE3 （BL21） and induced by IPTG. The Ferritin protein was successfully induced by IPTG, yielding a band that was consistent with the theoretical value （19.448 kD）. The survival rate of recombinant plasmid transferred E. coli was higher than that of empty vector transferred cells. Furthermore, the recombinant plasmid significantly improved the tolerance of E. coli to salinity stress. Our results suggest that Ferritin improves the ability of E. coli to cope with salinity stress and so may also play a role in salinity adaptation in P trituberculatus.