| 编码日本血吸虫大陆株磷酸丙糖异构酶(TPI)抗原基因的克隆与测序 |
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| 引用本文： | 缪应新,刘述先.编码日本血吸虫大陆株磷酸丙糖异构酶(TPI)抗原基因的克隆与测序[J].中国寄生虫学与寄生虫病杂志,1997,15(6):349-352. |
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| 作者姓名： | 缪应新 刘述先 |
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| 作者单位： | **华东医院上海市老年医学研究所; 中国预防医学科学院寄生虫病研究所 |
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| 摘 要： | 目的 :获得编码日本血吸虫大陆株磷酸丙糖异构酶 ( Sjc TPI)的 c DNA基因片段 ,对其克隆与测序。方法 :以日本血吸虫大陆株成虫总 RNA为模板 ,经过逆转录 -聚合酶链 (式 )反应( RT- PCR)获得编码 Sjc TPI抗原的 c DNA片段 ,并克隆于载体 M13mp18、M13mp19,双脱氧链末端终止法测 DNA序列。结果 :体外 PCR扩增获得了编码 Sjc TPI的 0 .75kb c DNA片段 ,6个重组克隆子经酶切鉴定均有 0 .75kb片段的插入。测得编码 Sjc TPI的 DNA序列。结论 :用自行设计的引物成功地扩增了编码日本血吸虫大陆株 TPI抗原的基因片段并进行了克隆、测序 ,为制备重组 TPI进行疫苗试验打下基础。
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| 关 键 词： | 日本血吸虫大陆株 磷酸丙糖异构酶 RT-PCR 基因克隆 DNA序列 |
CLONING AND SEQUENCING OF THE GENE ENCODING TPI ANTIGEN FROM CHINESE STRAIN OF SCHISTOSOMA JAPONICUM |
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| Miao Yingxin,Liu Shuxian.CLONING AND SEQUENCING OF THE GENE ENCODING TPI ANTIGEN FROM CHINESE STRAIN OF SCHISTOSOMA JAPONICUM[J].Chinese Journal of Parasitology and Parasitic Diseases,1997,15(6):349-352. |
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| Authors: | Miao Yingxin Liu Shuxian |
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| Affiliation: | **Shanghai Geriatrics Institute, Shanghai 200040
Institute of Parasitic Diseases; Chinese Academy of Preventive Medicine; Shanghai2 0 0 0 2 5 |
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| Abstract: | AIM:Cloning and sequencing the gene encoding TPI antigen from Chinese strain of S. japonicum. METHODS:RT- PCR technique was used to gain the target gene. After digestion with restriction enzyme, the gene was respectively cloned into the vectors M13mp18, M13mp19, then sequenced by the method of dideoxy- mediated chain- termination. RESULTS:The 0.75 kb c DNA encoding Sjc TPI antigen was amplified in vitro by RT- PCR using total RNA as a template. 0.75 kb insert DNA was identified in six recombinant clones by restriction enzymemapping. The DNA sequence of SjcTPI was determined. CONCLUSION: The process have been made succesfully in the cloning and sequencing the gene encoding SjcTPI. It will be scheduled to go ahead for vaccine trial probably using recombinant TPI antigen of S.japonicum. |
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| Keywords: | Schistosoma japonicum （Chinese strain） TPI RT- PCR gene clone DNA sequenc |
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