橡胶树HbCCoAOMT家族基因的鉴定

Genome-wide analysis of HbCCoAOMT gene family in rubber tree

为了进一步了解HbCCoAOMT基因在橡胶树(Hevea brasiliensis)生长发育和抵抗逆境中的作用,本研究通过分析巴西橡胶树的全基因组测序数据,对HbCCoAOMT家族基因进行鉴定分析,得到了40个HbCCoAOMT家族成员,探讨其在染色体上的分布、结构域特征及组织特异性。结果显示,橡胶树共有18条染色体,HbCCoAOMT成员只分布于1、3、4、11、13、17、18这7条染色体上。表达模式分析发现,有12个成员在所有5个组织中均有表达;HbCCoAOMT酶活检测结果表明,在橡胶树的茎中HbCCoAOMT酶活性最高。此外,从橡胶树中成功克隆获得表达丰度最高的HbCCoAOMT1基因,该基因CDS长746 bp,编码241个氨基酸。将HbCCoAOMT1构建在原核表达载体pET28a上,并在大肠杆菌E. coli BL21(DE3)中成功表达HbCCoAOMT1重组蛋白。分析HbCCoAOMT1启动子区域发现多种激素或胁迫相关顺式作用元件。

To further understand the role of the HbCCoAOMT gene in the growth,development,and stress resistance of the rubber tree(Hevea brasiliensis),the HbCCoAOMT family genes were identified and analyzed by examining the whole genome sequencing data of H.brasiliensis.A total of 40 members of the HbCCoAOMT family were identified,and their distribution on chromosomes,domain characteristics,and tissue expression profiles were analyzed.The results revealed that HbCCoAOMT members were only distributed in 7 chromosomes:1,3,4,11,13,17,and 18 out of the18 chromosones in rubber trees.Expression pattern analysis indicated that 12 members were expressed in all the five tissues.The HbCCoAOMT enzyme activity assay showed the enzyme activity was highest in the stems of H.brasiliensis.Moreover,HbCCoAOMT1,which had the highest expression abundance,was successfully cloned from H.brasiliensis.The CDS length of HbCCoAOMT1 is 746 bp,encoding 241 amino acids.HbCCoAOMT1 was inserted into the prokaryotic expression vector pET28a,and the recombinant HbCCoAOMT1 protein was successfully expressed in E.coli BL21(DE3).Analysis of the HbCCoAOMT1 promoter region showed that HbCCoAOMT1 contains various hormone-and stress-related cisacting elements.