C2株蓝氏贾第鞭毛虫cDNA文库的构建#br#

doi:10.3969/j.issn.10073205.2018.01.002

河北医科大学学报

C2株蓝氏贾第鞭毛虫cDNA文库的构建#br#

华北理工大学基础医学院病原生物学与免疫学系，河北唐山 063210

出版日期:2018-01-25 发布日期:2018-01-05
作者简介:刘绍伟(1987-)，男，安徽利辛人，华北理工大学基础医学院医学硕士研究生，从事病原生物学研究。〖
基金资助:
国家自然科学基金（31471954）；河北省自然科学基金（H2017209143）

Construction of C2 strain of giardia lamblia cDNA library

Department of Pathogenic Biology and Immunology, College of Basic Medicine, North China University of Science and Technology, Hebei Province， Tangshan 063210， China

Online:2018-01-25 Published:2018-01-05

摘要/Abstract

摘要： ［摘要］〓
〖HTH〗目的〖HTSS〗〖KG*2〗构建C2株蓝氏贾第鞭毛虫cDNA文库。
〖HTH〗方法〖HTSS〗〖KG*2〗Trizol对C2株贾第虫滋养体RNA进行提取，逆转录获得cDNA单链，LDPCR扩增获得ds cDNA，经纯化后，与pGADT7Rec共转化酵母株Y187中。通过营养缺陷型平板SD/Leu培养获得转化子即包含蓝氏贾第鞭毛虫cDNA文库，并对文库的库容及DNA重组率进行鉴定。
〖HTH〗结果〖HTSS〗〖KG*2〗以RNA为模板，经逆转录、LDPCR、纯化等，得到了较为理想的双链cDNA。将ds cDNA和pGADT7Rec转化到酵母菌株，经分离培养，最终得到转化成功的酵母转化子，建立了贾第虫的cDNA文库，文库容量为2.715×107/mL，重组率76.7%，插入片段的大小主要集中在300~1 000。
〖HTH〗结论〖HTSS〗〖KG*2〗成功的构建了较为理想的C2株贾第虫cDNA文库。

关键词: 贾第虫属, 基因文库, 双杂交系统技术

Abstract: ［Abstract］〓Objective〖HTSS〗〓To construct a cDNA Library of Giardia lamblia (C2) strains.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓By extraction of C2 strains of Giardia lamblia trophozoites RNA using the Trizol method, and by using RNA as a template by reverse transcriptase cDNA single strand. dscDNA was obtained by LDPCR amplification. dscDNA was purified and co transformed with pGADT7Rec in yeast strain Y187. Through auxotrophic medium SD/Leu screening, transformants including Giardia lamblia cDNA library were obtained. Then the storage capacity and DNA recombination rate of the library were identified.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓Using RNA as template, the ideal double stranded cDNA was obtained by reverse transcription, LDPCR, purification and other steps. The dscDNA and pGADT7Rec were transformed into yeast strain, and the transformants were successfully isolated and cultured. Finally, a cDNA Library of Giardia was constructed. The library capacity was 2.715×107/mL, the recombination rate was 76.7%, and the size of the inserted fragments was mainly between 300bp and 1000bp.
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓An ideal C2 Library of Giardia lamblia cDNA was constructed.

Key words: Giardia, gene library, twohybrid system techniques

刘绍伟，王〓洋，田喜凤. C2株蓝氏贾第鞭毛虫cDNA文库的构建#br#[J]. 河北医科大学学报, doi: 10.3969/j.issn.10073205.2018.01.002.

LIU Shaowei， WANG Yang， TIAN Xifeng. Construction of C2 strain of giardia lamblia cDNA library[J]. Journal of Hebei Medical University, doi: 10.3969/j.issn.10073205.2018.01.002.

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C2株蓝氏贾第鞭毛虫cDNA文库的构建#br#

Construction of C2 strain of giardia lamblia cDNA library

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