The gene structure and promoter region of the vaccine target aminopeptidase H11 from the blood-sucking nematode parasite of ruminants, Haemonchus contortus
The gene structure and promoter region of the vaccine target aminopeptidase H11 from the blood-sucking nematode parasite of ruminants, Haemonchus contortus
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反刍动物捻转血矛线虫氨肽酶H11疫苗基因结构及启动子区
DOI：
10.1007/s10142-010-0172-5
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发表时间：
2010-05
期刊：
影响因子：
--
通讯作者：
H.-L. Zhang
中科院分区：
文献类型：
--
作者：
X.-L. Jiang;A.-F. Du;Q.-J. Zhou;H.-L. Zhang
氨基肽酶H11是一种完整的膜糖蛋白，只存在于缠绕血圆线虫的肠道中，可使粪卵减少90%，而在大肠杆菌中表达的重组蛋白诱导的保护作用很小。为了进一步研究H11基因的特征，我们通过逆转录聚合酶链式反应扩增了H11基因的mRNA，然后分离了其1,517个碱基的5‘-侧翼区，并对其基因组结构进行了测定。H11基因由24个内含子隔开的25个外显子组成，基因组DNA全长14,959个碱基。对H11基因5‘侧翼区1,517个碱基对的分析表明，可能存在一个“TATA缺失”启动子。H11亚型H11-4的最后一个外显子及其3‘-非编码区的部分序列也位于H11基因上游，具有相同的转录方向。将H11基因5‘侧翼区1,517个碱基和第一外显子部分亚克隆到绿色荧光蛋白报告基因上游的载体上，显微注射入秀丽线虫的性腺中。转化的动物在L4幼虫期和成虫的远端肠道均有荧光。本研究描述了氨基肽酶H11的基因结构，展示了其启动子区域在自由生活线虫和吸血线虫之间的不同转录模式，并强调了线虫作为异源系统在研究H11亚型生物学作用方面的应用。
Aminopeptidase H11, an integral membrane glycoprotein present only in the gut of Haemonchus contortus, could provide substantial protection as shown by 90% reduction in fecal egg counts, while its recombinant version expressed in E. coli induced little. To investigate the characteristics further, we amplified mRNA of H11 gene via reverse transcriptase polymerase chain reaction, followed by isolation of its 1,517-bp 5'-flanking region and determination of its genomic organization. The H11 gene contained 25 exons separated by 24 introns and spans 14,959 bp of genomic DNA. Analysis of the 1,517 bp 5'-flanking region of the H11 gene revealed a putative "TATA-less" promoter. Partial sequences of the last exon and its 3'-UTR of H11 isoform H11-4 were also identified upstream to the H11 gene with the same transcription orientation. The 1,517-bp 5'-flanking region and part of the first exon of the H11 gene were subcloned into the vector upstream of green fluorescence protein reporter gene and microinjected into the gonads of Caenorhabditis elegans. The transformed animals exhibited fluorescence in the distal intestine in the L4 larvae stage and adult worms. This study characterized gene structure of aminopeptidase H11, demonstrated different transcriptional pattern of its promoter region between free-living and blood-sucking nematode species, and highlights the utility of C. elegans as a heterologous system to study the biology roles of H11 isoforms.
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影响因子：
2.2
作者：
L. Vervelde;M. V. van Leeuwen;M. Kruidenier;F. Kooyman;J. Huntley;I. van Die;A. Cornelissen
通讯作者：
L. Vervelde;M. V. van Leeuwen;M. Kruidenier;F. Kooyman;J. Huntley;I. van Die;A. Cornelissen
DOI：
10.2307/3284267
发表时间：
1997-10
期刊：
The Journal of parasitology
影响因子：
--
作者：
M. L. Rhoads;R. Fetterer;J. Urban
通讯作者：
M. L. Rhoads;R. Fetterer;J. Urban
影响因子：
1.6
作者：
E. Masler
通讯作者：
E. Masler
DOI：
10.1074/jbc.m109.026948
发表时间：
2009-09
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
K. Ohneda;S. Ohmori;Y. Ishijima;Mayu A Nakano;Masayuki Yamamoto
通讯作者：
K. Ohneda;S. Ohmori;Y. Ishijima;Mayu A Nakano;Masayuki Yamamoto
影响因子：
1.3
作者：
M. Ep
通讯作者：
M. Ep