水生生物学报2011，Vol.35Issue(4)：672-680,9.DOI:10.3724/SP.J.1035.2011.00672
湖北钉螺肝胰腺cDNA文库的构建和序列分析
CONSTRUCTION AND ANALYSIS ON CDNA LIBRARY OF ONCOMELANIA HUPENSIS HEPATOPANCREAS
摘要
Abstract
The snail Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum which causes schistosomiasis endemic in the Fast East, especially in mainland China. After miracidium penetrated into the snail host, the schistosome larvae develop into sporocyst, move to snail hepatopancreas and develop into cercariae at last, which is the infection stage for the final host including human. Hepatopancreas is not only the necessary organ for development of schistosome larvae in snail host, but also be rich of hemocyte, which was thought to mediate the defense system of O. Hupensis in response to schistosome infection. The present study was thus designed to construct a cDNA library of O. Hupensis hepatopancreas, and give some information to deeply understand the interaction between the snail host and schistosome parasite through obtained genomic data. Total RNA were extracted from O. Hupensis hepatopancreas and mRNA were purified by using Poly Attract mRNA Isolation Systems IV. The cDNA library was then constructed from purified mRNA by using CloneMiner? cDNA Library Construction Kit under the manual direction. Totally, library had 5.6x104cfu/mL in initial liter, and 6.72xl05cfu/mL in capacity. The percentage of recombination was about 93%, with the length of inserts being 400-2700 bp. 1728 positive clones were sequenced, and a total of 1520 were successfullysequenced with the yield of 1393 expressed sequence tags (ESTs) longer than 100 bp. Through initial gene splicing, 698 unigenes were formed from 1393 ESTs, including 234 contigs and 464 singletons. After BLAST (Basic Local Alignment Search Tool) analysis of these sequences, 111 (15.90%) unigenes were assigned to have homology with some genes in the GenBank, which could be divided into four types, including Cellular Processes and Signaling (21.62%), Information Storage and Processing (46.85%), Metabolism (27.93%) and Poorly Characterized (3.60%) based on the protein function through COG analysis. Other 587 (84.10%) unigenes had no homology with any genes in present GenBank data, which might be potential for further novel gene discovery. O. Hupensis hepatopancreas cDNA library have been constructed, with plenty of new genes have been discovered, which supply genomic platform for further study. In addition, some immunology related genes such as glutathione S-transferase, chitinase, ferritin-like protein, glutathione peroxidase and heat shock protein family have been found, which may indicate that the hepatopancreas participates in the immunology activity of snail against parasite. In summary, this study provides plenty of genomic data of O. Hupensis, which can supply some information for further investigations such as discovery of new immunology of snail host, and interaction between O. Hupensis and S. Japonicum.关键词
湖北钉螺；肝胰腺；cDNA文库/ESTKey words
Oncomelania hupensis/ Hepatopancreas/ cDNA library/ Expressed sequence tag分类
生物科学引用本文复制引用
朱丹丹,赵琴平,聂品..湖北钉螺肝胰腺cDNA文库的构建和序列分析[J].水生生物学报,2011,35(4):672-680,9.基金项目
中国科学院知识创新工作重要方向性项目(KSCX2-YW-N-055,KSCX2-W-2-0808)；国家自然科学基金(30770353)资助 （KSCX2-YW-N-055,KSCX2-W-2-0808）
