药用植物黄花蒿ATP合成酶电子克隆及生物信息学分析

In Silico Cloning and Bioinformatics Analysis of Medicinal Plant Artemisia annua ATP Synthetase

本文通过电子克隆的方法获得黄花蒿ATP合成酶的基因完整序列并对该蛋白特性进行相应分析.以Accession number KJ434435.1为探针,对黄花蒿的EST数据库进行搜索,获得同源较高的序列,用相关生物软件DNAMAN、MEGA进行拼接组装,并对获得核苷酸和氨基酸进行生物信息学分析.结果发现获得黄花蒿ATP合成酶基因拼接群1963bp,含有一个完整的开放阅读框(ORF)序列(1479bp),编码492个氨基酸,该蛋白质含15个α螺旋,23个β折叠及27个无规则卷曲.黄花蒿ATP合成酶蛋白的分子质量为52830.4,分子式为C_(2342)H_(3778)N_(636)O_(721)S_(14),理论等电点为5.18,该蛋白的GRAVY值为-0.048,具有亲水性的水溶性蛋白.该蛋白质序列有丝氨酸磷酸化位点(Ser)9个、苏氨酸磷酸化位点(Thr)6个、酪氨酸磷酸化位点(Tyr)4个.通过分析获得黄花蒿ATP合成酶完整的c DNA序列,该氨基酸序列不存在信号肽,无跨膜现象、非分泌型蛋白,为黄花蒿ATP合成酶实验室研究提供一定的理论基础.

[ Objective ] Used the in silico cloning method to get ATP synthase whole cDNA sequence of Artemisia annua and analyzed the protein characteristics. [ Method ] Used GenBank accession number KJ434435.1 as probe to search some high homology sequences in Artemisia annua EST data-base, and used biology software DNAMAN, MEGA to join these sequences together and assembly, then bioinformatics analysis of the nucleotide, amino acid sequences. [ Results] Obtained a ATP synthase gene cDNA sequence,and the in silico cloned contig cDNA is 1963 base pairs long with a complete open reading frame（ORF） containing 1479 nucleotides encoding a protein of 492 amino acid. The protein contains 15 alpha helix, 23 beta folding and 27 random curl. Artemisia annua ATP synthetase protein molecular weight is 52830.4, its molecular formula is C2342H6360721 S14 , theoretical isoelectric point 5.18, the GRAVY value of the protein-0. 048, and it is a hydrophilic water-sol-uble protein. Artemisia annua ATP synthetase has 9 serine phosphorylation sites, 6 threonine phos-phorylation sites and 4 tyrosine phosphorylation sites. [ Conclusion ] The whole Arternisia annua ATP synthetase cDNA sequence is succeed to gain by in silico cloning method. The amino acid series have no signal peptide, transmembrane phenomenon and non-secretory proteins. It will offer theory and preparative to clone Artemisia annua ATP synthetase in laboratory.