巴西橡胶树HbCPT7基因的克隆与表达分析

摘要

cis-prenyltransferase is a key enzyme for natural rubber biosynthesis in rubber trees.A novel gene encoding cis-prenyltransferase was isolated by RT-PCR from the latex of rubber tree clone Reyan7-33-97 and designated as HbCPT7.Bioinformatic analysis showed that the predicted protein contained 296 aa and harbored five typical conserved motifs of the cis-prenyltransferases from other plants.Tapping as well as methyl jasmonate application up-regulated the expression of HbCPT7 gene,suggesting that HbCPT7 played an important role in natural rubber regeneration after tapping and might be a potential target of jasmonate signaling.SDS-PAGE and western blotting showed that the recombinant HbCPT7-his protein was successfully expressed in Escherichia coli after IPTG induction.These results lay a foundation for elucidation the roles of HbCPT7 in natural rubber biosynthesis.%来自巴西橡胶树的天然橡胶主要成份是顺式聚异戊二烯高分子化合物,顺式异戊二烯转移酶是天然橡胶生物合成中的关键酶之一.利用RT-PCR技术从巴西橡胶树胶乳中克隆出一个新的顺式异戊二烯转移酶基因HbCPT7.生物信息学分析表明该基因编码296个氨基酸,含有顺式异戊烯基转移酶典型的5个高度保守结构域.qRT-PCR分析表明该基因分别受割胶和茉莉酸甲酯上调诱导表达,推测该基因在割胶后胶乳再生中发挥重要的作用,亦可能是茉莉酸信号调控的靶标.SDS-PAGE和western blotting分析表明重组HbCPT7-His蛋白在大肠杆菌中成功由IPTG诱导表达.这些结果为进一步研究HbCPT7基因功能打下良好基础.