花生AhSOS2基因原核表达载体的构建及表达

Construction and Expression of Prokaryotic Expression Vector of AhSOS2 Gene from Peanut

SOS2(Salt Overly Sensitive 2)是植物中一类耐盐相关基因的统称,在植物对盐害的响应及适应过程中具有重要作用。本试验以花生AhSOS2基因全长cDNA为模板,经PCR扩增获得AhSOS2的蛋白编码区,将该区段与表达载体pET-28a(+)融合,构建获得原核表达载体pET-28a-AhSOS2,并进一步在大肠杆菌BL21(DE3)中进行表达。经1.0 mmol/L IPTG诱导后,获得一条约51 kD的融合蛋白条带,且随着IPTG诱导时间的延长,蛋白表达量逐渐提高,诱导8 h可获得较高的蛋白表达水平。

SOS2 （ Salt Overly Sensitive 2 ） is a kind of salt-tolerant related genes in plant .It plays an im-portant role in the response and adaptation process of plants to salt damage .Using the full-length cDNA of AhSOS2 gene as template , its coding region was amplified by PCR method .The recombined prokaryotic ex-pression vector pET-28a-AhSOS2 was constructed successfully by fusing AhSOS2 with expression vector pET -28a（＋）.Then the recombinant was expressed in E.coli strain BL21（DE3）.Inducing with 1.0 mmol/L IPTG, a 51 kD fusion protein was obtained .Its expression quantity improved gradually with the extension of induction time, and attained the highest level after inducing for 8 hours.