作者：
Yan Liu;Qingren Zeng;Jun Zhou;Shenghui Yang（杨胜辉）;Biyuan Liu（刘碧源）;...
作者机构：
Department of Parasitology,School of Medicine,University of South China;Center of cell and Molecula;Center of cell and Molecular Biology Experiment,Xiangya School of Medicine,Central South University;Experimental Center of Xiangya Third Hospital,CSU;Department of Pathogenic Biology and Immunology,Hunan University of Chinese Medicine
作者：
Yang, S. H.（杨胜辉）;Brindley, P. J.;Zeng, Q. R.*;Zeng, T. B.;Li, Y. S.;...
作者机构：
[Liu, Y.; Yang, S. H.; Liu, B. Y.; Li, Y. S.; Cai, L. T.; Zeng, T. B.; Lan, L. M.; Zeng, Q. R.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.;[Yang, S. H.; Wu, C. R.; Tan, Z. J.; Lu, F. G.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.;[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.;[Liu, Y.; Liu, B. Y.; Zeng, T. B.] Univ South China, Sch Med, Hengyang, Peoples R China.;[Li, Y. S.; McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.
会议名称：
12th International Congress of Parasitology (ICOPA)/6th Novel Approaches to the Control of Helminth Parasites of Liverstock Conference
会议时间：
AUG 15-20, 2010
会议地点：
Melbourne, AUSTRALIA
会议主办单位：
[Yang, S. H.;Zeng, Q. R.;Zeng, T. B.;Li, Y. S.;Liu, Y.;Liu, B. Y.;Cai, L. T.;Lan, L. M.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.^[Yang, S. H.;Wu, C. R.;Lu, F. G.;Tan, Z. J.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.^[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.^[Zeng, T. B.;Liu, Y.;Liu, B. Y.] Univ South China, Sch Med, Hengyang, Peoples R China.^[Li, Y. S.;McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.^[Zhou, J.] Cent S Univ, Expt Ctr Xiangya Third Hosp, Changsha, Hunan, Peoples R China.
摘要：
Retroviral transduction of cultured schistosomes was explored in our study. The vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter gene under the control of retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce Schistosoma japonicum (Sj) to investigate the utility of retrovirus mediated Sj transgenesis and to investigate the activity as human telomerase as a reporter transgene in schistosomes. The presence of transgene hTERT was detected by PCR. Furthermore, analysis of RNAs from transduced parasites, immunohistochemistry of thin sections and immunoblot analysis revealed expression of hTERT transgene in the transduced worms. These findings indicated that Sj could be effectively transduced by VSVG pseudotyped retrovirus carrying the hTERT gene.
摘要：
Although draft genome sequences of two of the major human schistosomes, Schistosoma japonicum and Schistosoma mansoni are available, the structures and characteristics of most genes and the influence of exogenous genes on the metabolism of schistosomes remain uncharacterized. Furthermore, which functional genomics approaches will be tractable for schistosomes are not yet apparent. Here, the vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter, under the control of the retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce S. japonicum to investigate the utility of retrovirus-mediated transgenesis of S japonicum and the activity of human telomerase reverse transcriptase as a reporter transgene in schistosomes. Schistosomules perfused from experimentally infected rabbits were cultured for 6 days after exposure to the virions after which genomic DNAs from virus exposed and control worms were extracted. Analysis of RNA from transduced parasites and immunohistochemistry of thin parasite sections revealed expression of hTERT in the transduced worms. Expression of hTERT was also confirmed by immunoblot analysis. These findings indicated that S. japonicum could be effectively transduced by VSVG-pseudotyped retrovirus carrying the hTERT gene. Given the potential of hTERT to aid in derivation of immortalized cells, these findings suggest that this pantropic retroviral approach can be employed to transduce cells from specific tissues and organs of schistosomes to investigate the influence of transgene hTERT on growth and proliferation of schistosome cells. (C) 2010 Elsevier B.V. All rights reserved.
