Clone of Full-length cDNA Sequence of Actin Gene of Eleutherococcus Senticosus and Bioinformatics Analysis
10.11842/wst.2013.09.007
- VernacularTitle:刺五加Actin基因cDNA全长序列的克隆与生物信息学分析
- Author:
Lihua CHAI
;
Leshan XIU
;
Mi ZHOU
;
Zhaobin XING
- Publication Type:Journal Article
- Keywords:
Eleutherococcus senticosus;
Actin;
clone;
bioinformatics analysis
- From:
World Science and Technology-Modernization of Traditional Chinese Medicine
2013;(9):1901-1906
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to clone the full-length cDNA sequence of actin gene of Eleutherococcus sentico-sus. And bioinformatics analysis was used. The total RNA was isolated from leaves of E. senticosus , and cDNA was synthesized by reverse transcription of total RNA. Primers were designed according to the conserved se-quence that had been cloned of Actin of E. senticosus . Then, the 3'and 5' cDNA fragments were cloned by nested PCR . The full-length gene was obtained by gene splicing method . Sequencing results were compared and treated with similarity analysis by blast analysis in the GenBank. Protein secondary structure and tertiary struc-ture of Actin of E. senticosus was predicted by online software. The results showed that the full-length cDNA of Actin of E. senticosus is 1507 bp, which named EsActin1, GenBank accession No. KC469585. The conserved sequence, which contained a 1134 bp open reading frame that encoding a 377 amino acid residues, a 5'-UTR of 140 bp and a 3'-UTR of 233 bp. Homologous alignment showed that it shared over 75% nucleotide se-quence similarity and over 94% amino acid sequences similarity with Actins in other plants. It was concluded that this study first isolated and reported the full-length cDNA sequence of actin gene of E. senticosus , and laid a foundation for the molecular biology research of E. senticosus .