【摘要】 重金属关联异戊二烯化植物蛋白(Heavy metal-associated isoprenylated plant proteins,HIPPs)由于其独特的重金属结合域(heavy metal associated domains,HMA)和异戊二烯序列(isoprenylation motif)的结构特点,使其成为一类重要的金属分子伴侣(Metallochaperones)。本研究利用RT-PCR的方法,从甜菜体内克隆获得了Beta vulgaris heavy metal-associated isoprenylated plant protein24(BvHIPP24)基因。该基因CDS全长为444 bp,编码了147个氨基酸,分子量为16.38 kDa。利用大肠杆菌表达载体构建pEASY-Blunt E1-BvHIPP24重组质粒,并转化到BL21中。通过1 mmol/L IPTG诱导发现:在0.5 mmol/L Cd2+逆境胁迫下,表达BvHIPP24重组菌的生长趋势要明显优于对照菌株,并具有更高的Cd耐受性。因此可以推断,甜菜BvHIPP24基因在镉逆境胁迫过程中发挥着重要的作用,并有可能参与到甜菜细胞重金属离子吸收、转运、区隔以及代谢平衡调控过程中。更多还原

【Abstract】 Because of unique heavy metal associated domains(HMA) and isoprenylation motif, heavy metalassociated isoprenylated plant proteins(HIPPs) are important members in Metallochaperones. In this study, Beta vulgaris heavy metal-associated isoprenylated plant protein24(BvHIPP24) was cloned by RT-PCR in sugar beet. Its full length of CDS was 444 bp, encoding 147 amino acids with a molecular weight of 16.38 kDa. The recombinant plasmid of pEASY-Blunt E1-BvHIPP24 was constructed and transformed into BL21. The results showed that, by 1 mmol/L IPTG induction, reconstructed E. coli expressing BvHIPP24 with superior Cd tolerance grew better than the control under 0.5 mmol/L CdCl2 stress. Hence, it could be concluded that BvHIPP24 played a vital role in cadmium stress process in sugar beet, and might be involved in cell heavy metal ion absorption, transportation,compartmentalization and metabolic homeostasis.更多还原