[1]胡晓飞,王英,潘春江,等.大劣按蚊TEP1 cDNA的克隆和生物信息学分析[J].陆军军医大学学报(原第三军医大学学报),2009,31(11):993-996.
 HU Xiao-fei,WANG Ying,PAN Chun-jiang,et al.Cloning and bioinformatic analysis of cDNA fragment of TEP1 gene of Anopheles dirus[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(11):993-996.
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大劣按蚊TEP1 cDNA的克隆和生物信息学分析(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第11期
页码:
993-996
栏目:
论著
出版日期:
2009-06-15

文章信息/Info

Title:
Cloning and bioinformatic analysis of cDNA fragment of TEP1 gene of Anopheles dirus
作者:
胡晓飞王英潘春江刘涛谢佳何谐姜晓梅黄复生
第三军医大学:学员旅7队,基础医学部病原生物学教研室,学员旅20队,学员旅21队
Author(s):
HU Xiao-fei WANG Ying PAN Chun-jiang LIU Tao XIE Jia HE Xie JIANG Xiao-mei HUANG Fu-sheng
7th Student Team; Department of Pathogenic Biology, College of Basic Medical University, 20th Student Team, 21st Student Team, Third Military Medical University, Chongqing 400038, China
关键词:
大劣按蚊TEP1RT-PCR生物信息学天然免疫
Keywords:
Anopheles dirus thioester-containing protein 1 RT-PCR bioinformatics innate immunity
分类号:
R318.04;R384.1;R394.3
文献标志码:
A
摘要:
目的   克隆大劣按蚊(Anopheles dirus)含硫酯键蛋白1(thioester-containing protein 1,TEP1)的cDNA,并分析其序列。   方法   根据已报道的冈比亚按蚊等多种昆虫的TEP1氨基酸保守序列设计简并引物,以大劣按蚊血淋巴细胞总RNA为模板,进行RT-PCR扩增。对目的片段纯化回收,经TA克隆后测定其核苷酸序列。利用DNASIS程序分析该序列,并推导其氨基酸序列,最后利用BLAST和DNASIS等进行不同昆虫TEP1氨基酸序列的比对。   结果   RT-PCR扩增出一约770 bp的条带,TA克隆后DNA测序发现,阳性克隆的核苷酸序列长774 bp,推导其氨基酸序列长258 aa。生物信息学分析表明,该序列与冈比亚按蚊、阿拉伯按蚊、斯氏按蚊TEP1基因的氨基酸序列同源性分别达72%、72%和59%,与冈比亚按蚊的TEP4氨基酸序列同源性为59%。   结论   从大劣按蚊血淋巴细胞中成功克隆出了TEP1的cDNA部分序列。
Abstract:
Objective   To clone the cDNA fragment of thioester-containing protein 1 (TEP1) of Anopheles dirus and analyze the sequence with bioinformatic methods.    Methods   The target cDNA fragment was amplified by RT-PCR with degenerated primers designated based on the conserved amino acid sequences of TEP1 of Anophelesgambiae and other insects. And total RNA from haemocytes of Anopheles dirus was used as the template of RT-PCR. The PCR product was purified and T/A cloned, followed by sequencing the positive clone. Then the sequence was analyzed with DNASIS software and the amino acid sequence was deduced. The obtained amino acid sequence was aligned with TEP1 amino acid sequences of the other insects with BLAST and DNASIS.     Results   A band of approximately 770 bp was amplified by RT-PCR. A nucleotide acid sequence with 774 bp in length was obtained by T/A cloning and sequencing. The deduced amino acid sequence was 258 aa in length, and had a sequence homology of 72%, 72% and 59%  respectively when compared with the TEP1 sequences of Anopheles gambiae, Anopheles arabiensis and Anopheles stephensi.    Conclusion   The cDNA fragment of TEP1 is successfully cloned from the haemocytes of Anopheles dirus, which provides a foundation for further study on function of the molecular.

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更新日期/Last Update: 2009-05-21