Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia  

作　　者：Jun-Li Guo Jie Jiang Wen-Yu Zheng Ming-Luan Li Xi-Feng Tian Xian-Min Feng Yue-Hua Wang Xiao-Hong Ju Yue-Qiong Kong 

机构地区：[1]School of Laboratory Medicine,Jilin Medical College [2]Hainan Provincial Key Laboratory of Tropical Medicine,Hainan Medical College [3]Department of Hand Microsurgery,Central Hospital of Jilin City [4]College of Life Science,Hebei united university

出　　处：《Asian Pacific Journal of Tropical Medicine》2012年第12期931-934,共4页亚太热带医药杂志（英文版）

基　　金：supported by grants from the Education Department of Jilin Province(2010D532);the open grant from the State Key Laboratory of Genetic Resources and Evolution,Kunming Institute of Zoology(GREKF08-07);Natural Grant of Jilin City(201032243);Natural Foundation of Hainan Province of China(310043 and 811197);Key Project of Hainan Provincial Bureau of Health(No2010-41)

摘　　要：Objective:To construct rapidly a full-length cDNA library from nanogram amounts total RIMA of Giardia lamblia(G.lamblia) trophozoites stocked in RNA stabilization reagent.Methods:Total RNA of Giardia was extracted using Trizol reagent.A full-length cDNA library of G.lamblia trophozoites was constructed by a long-distance PCR(LD-PCR) method.The recombinant rate and the coverage rate of full-length clones of the library were evaluated.The inserted fragments were identified and sequenced by PCR amplification.Results:The titer of cDNA library was 3.85×10~7 pfu/mL.The length of inserted fragments ranged from 0.4 to 2.5 kb,and the recombination efficiency accounted for 100%(20/20).The coverage rate of full-length clones is high(17/20). Conclusions:The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature.The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA.Objective:To construct rapidly a full-length cDNA library from nanogram amounts total RIMA of Giardia lamblia(G.lamblia) trophozoites stocked in RNA stabilization reagent.Methods:Total RNA of Giardia was extracted using Trizol reagent.A full-length cDNA library of G.lamblia trophozoites was constructed by a long-distance PCR(LD-PCR) method.The recombinant rate and the coverage rate of full-length clones of the library were evaluated.The inserted fragments were identified and sequenced by PCR amplification.Results:The titer of cDNA library was 3.85×10~7 pfu/mL.The length of inserted fragments ranged from 0.4 to 2.5 kb,and the recombination efficiency accounted for 100%(20/20).The coverage rate of full-length clones is high(17/20). Conclusions:The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature.The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA.

关 键 词：GIARDIA LAMBLIA CDNA LIBRARY RNA stabilization REAGENT 

分 类 号：R382.2[医药卫生—医学寄生虫学]