摘要
本研究根据其它植物actin基因的保守序列设计一对简并性引物,以3周龄梭梭整株总RNA为模板,采用RT-PCR的方法扩增出actin基因片段并克隆到PUCm-T载体,阳性克隆经PCR检测后进行测序。序列分析结果表明:该片段长约598 bp,编码198个氨基酸;该序列与GenBank中现有的植物Actin比对发现,同源性均高达84%,同时,翻译得到的氨基酸序列与其它植物的氨基酸序列同源性高达99%。表达实验分析表明在不同胁迫条件下该基因在梭梭不同组织中表达量恒定,可作为内参基因。梭梭actin基因片段的克隆可为研究重要抗逆功能基因在梭梭中的表达和调控机制奠定基础。
In this paper degenerated primers were designed according to the conserved sequences of the actin gene from other plant species. Total RNA was extracted from the whole plant of Haloxylon ammodendron. actin gene fragment was obtained by reverse transcription polymerase chain reaction（RT-PCR） and cloned into PUCm-T vector. The positive clone identified by PCR was sequenced. The sequenced results indicated that the actin gene fragment of H. ammodendron contains about 600 bp, encoding 198 amino acids. Homology comparison with actin genes from other plant species in the GenBank indicated that HaACT shared more than 84% nucleotide and 99%amino acids sequence homology. Expression pattern analysis showed that under various stress conditions the gene expressed stably in the different organization of H. ammodendron. Our result confirmed that HaACT can be used as a housekeeping gene. This work lays a foundation for application of actin gene in characterizing other important genes relevant to stress resistance in xerophyte H. ammodendron.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2014年第3期598-603,共6页
Genomics and Applied Biology
基金
教育部"长江学者和创新团队发展计划"(IRT13019)
"新世纪优秀人才支持计划"(NCET-11-0217)
国家自然科学基金项目(31222053)
中央高校基本科研业务费专项资金(lzujbky-2013-k08)共同资助
关键词
梭梭
肌动蛋白基因
克隆
序列分析
Haloxylon ammodendron
actin gene
Cloning
Sequence analysis
