玉米高光效基因ppdk的克隆及其生物信息学分析

目的]获得玉米(Zea mays)的丙酮酸磷酸双激酶基因(以下简称ppdk),并对其进行生物信息学分析.方法]使用Trizol提取玉米SC704的总RNA,并用特异引物对其进行RT-PCR扩增,将得到的cDNA片段连接到pGEM-T载体后转化大肠杆菌DH5α,然后对阳性克隆进行测序,并对序列进行生物信息学分析.结果]获得的玉米PPDK基因CDS全长2781bp,与玉米z561ppdk基因同源性为99;;其编码的多肽链包含926个氨基酸,与玉米PPDK同源性为97;.结论]克隆了玉米C4型丙酮酸磷酸双激酶(PPDK)基因,它所编码的氨基酸序列具备PPDK蛋白的保守序列和催化活性中心区域.该基因的成功克隆为今后利用其改造C3植物的光合效率以提高粮食单产奠定了良好的基础.

Cloning and Bioinformatics Analysis of High Photosynthetic Gene ppdk in Zea Mays

【Objective】To clone the Pyruvate Orthophosphate Dikinase(PPDK)gene from Zea mays and analyze it by Bioinformatics.【Method】Primarily,the total RNA was extracted from Zea mays by the Trizol extraction and the cDNA clone was constructed by RT-PCR amplified with the gene specific primers.Then positive clones were sequenced and analyzed by Bioinformatics.【Result】The electrophoresis showed that the length of Coding Sequence(CDS) of PPDK gene in zea mays was 2 781 bp and highly related to zea mays cultivar z561 with 99% of homology.Additionally,the polypeptide chains coded by ppdk included 926 amino acids,which were 97% homology with pyruvate orthophosphate dikinase basing on amino acids sequences comparison.【Conclusion】The PPDK gene in zea mays has been obtained and the amino acids sequence coded by the gene conserve and contain active catalyst central site of PPDK.The successful cloning of the gene has set the base for utilizing improvement of C3 plant photosynthetic efficiency to further increase unit area yield of grain.