The Construction of the Eukaryotic Expression Vector of Glycerophosphodiester Phosphodiesterase Gene from Treponema pallidum and its Expression in Hela Cells-中国期刊网

摘要 Objective:ToconstructtherecombinantplasmidcontainingGlycerophosphodiesterphosphodiesterase(Gpd)genefromTreponemapallidumandtransfectitintoHelacellstoexpresstheencodedoutermembraneprotein.Methods:TheGpdgenewasamplifiedfromthegenomicDNAofT.pallidumbypolymerasechainreaction(PCR)andinsertedintocloningvectorpUCm-T.TheinsertedGpdgenewassubclonedintotheappropriatesiteofpcDNA3.1(+)vector.Afteridentificationbysequencingandrestrictiveenzymesdigestion,therecombinantplasmidwastransfectedintoHelacellsusingliposomes.TheexpressedproteinwasidentifiedbyimmunocytochemistryandWesternblot.Results:ThetargetGpdgenesegmentwasapproximately1059bp.TheDNAsequenceoftheGpdgenecontainedinthepcDNA3.1(+)vectorwasconsistentwiththepublishednucleotidesequence.ThehomologyofthenucleotideandputativeaminoacidsequencesoftheGpdgenebetweenT.pallidumsubsp.pallidumNicholsandvariouspathogenictreponemalstrainsrangedfrom98%to100%.ImmunocytochemistryandWesternblotanalysisshowedthattheconstructedGpd-pcDNA3.1(+)vectorexpressedafusionproteinwithacalculatedmolecularmassof41KDainHelacellsandthattheexpressedproteinreactedwiththeserafromsyphilispatients.Conclusion:ThesuccessfulconstructionandexpressionoftheeukaryoticexpressionplasmidoftheGpdgenefromT.pallidumprovideapromisingtooltofurtherstudythebiologicalactivityofT.pallidumanddevelopaDNAvaccineforsyphilis.

1苏立;陈运贞;张晓刚;佘强. Construction of the Eukaryotic Expression Vector with EGFP and hVE GF121 Gene and its Expression in Rat Mesenchymal Stem Cells.心血管疾病,2005-01.
2赵飞骏;吴移谋;刘双全;余敏君. Construction of the Eukaryotic Expression Vector for Outer Membrane Protein Tp92 from Treponema pallidum and Its Preliminary Study on the Immune Responses in New Zealand Rabbits.基础医学,2004-03.
3FEI JUN ZHAO;YI Mou WU;XIAO HONG ZHANG;SnUANG QUAN LIU;MIN JUN YU;LI SHENG ZHAN. Eukaryotic expression of outer membrane protein Gpd from Treponema pallidum and preliminary studies on its immune response in rabbits.基础医学,2005-02.
4WEI HE;FANG LIU;LING BO LIU;MIN ZHANG;ZHONG BO HU;PING ZOU. Construction of the recombinant expression vector for CD80-IgG fusion gene and its expression in Chinese hamster ovary cells.基础医学,2005-04.
5XichunHan;HongZhang;MingkuJia;GangHan;WeidongJiang. Expression of TIMP-3 Gene by Construction of a Eukaryotic Cell Expression Vector and Its Role in Reduction of Metastasis in a Human Breast Cancer Cell Line.免疫学,2004-04.
6Wei Han Qin Gang Hu Zhi Yong Wang Xu Dong Yang Xiao Feng Huang. Construction and evaluation of eukaryotic expression vector expressing VEGF shRNA.口腔医学,2008-08.
7熊礼宽;周华;王慧;姜维娜;洪福昌;曾序春;罗斌. Cloning and Expression of the Tppl7 Gene of Treponema pallidum And Clinical Application.皮肤病学与性病学,2001-01.
8刘双全;吴移谋;赵飞骏;曾铁兵. Construction and Expression of Tp0453 Recombinant Protein of Treponema pallidum and Development of Indirect ELISA for Diagnosing Syphilis.皮肤病学与性病学,2005-01.
9WANG Chuan-ming CAI Xiao-tang SHEN Han-bin ZHANG Shao-yan LOU Chao-yang. Construction, Stable Expression of Survivin shRNA Vector.生物医学工程,2011-01.
10Yongjun Sui, Guangzhi Ding, Pei-Hsien Tang, YuweiFan, Ning MaoBeijing Institute of Basic Medical Sciences, Beijing100850. The cloning of HLA-DR gene and its expression in mouse melanoma cells.临床医学,1997-03.

The Construction of the Eukaryotic Expression Vector of Glycerophosphodiester Phosphodiesterase Gene from Treponema pallidum and its Expression in Hela Cells

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The Construction of the Eukaryotic Expression Vector of Glycerophosphodiester Phosphodiesterase Gene from Treponema pallidum and its Expression in Hela Cells

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