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Phenotypic Knockout of CXCR4 on Molt-4 with SDF-1α/54 Attached with KDEL
作者姓名:CHEN Hong-yuan TAN Yi GUO Zhi-gang MA Wei-feng CAI Shao-xi DU Jun HUANG Jun CAI Shao-hui
作者单位:[1]Key Laboratory for Biomechanics & Tissue Engineering of the State Ministry of Education, College of Bioengineering Chongqing University, Chongqing 400044, China [2]Department of Microbiology-Immunology School of Guangdong Pharmacy University, Guangzhou 510006, China [3]Department of Clinical Pharmacology, Pharmacy School of Jinan University, Guangzhou 510632, China [4]School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou 510612, China [5]School of Pharmaceutical Sciences, Wenzhou Medical College, Wenzhou 325025, China [6]Department of Microbiology-Immunology School of Sun Yat-Sen University, Guangzhou 510612, China
基金项目:National Natural Science Foundation of China;Grant number:30572209
摘    要:
Objective : To investigate the mechanism of phenotypic knockout of CXCR4 on T-cell leukemia cell line Molt-4 via SDF-1α/54/KDEL intrakine technology, which the mutant SDF-1α/54, human stromal cell-derived Faceor-1 (SDF-1α) was deleted its C- terminal α-helix and attached with a endoplasimc reticulum retention signal 4-peptide- KDEL encoding gene, so that retain the newly synthesized receptor CXCR4 within the Molt-4 cells endoplasmic reticulum. Methods: The recombinant vector pEGFP-C3/SDF- 1α/54/KDEL were transfected into Cos-7 cells by liposome, SDF-1α/54/KDEL fusion protein was confirmed with western blot. The recombinant plasmids were transfected transiently into Molt-4 by electroporation. Results:Western blot confirmed SDF-1α/54/KDEL expression in Cos-7. A dramatic downregulation of CXCR4 expression on Molt-4 was demonstrated by flow cytometric (FCM) analysis. Conelusion:SDF-1α/54/KDEL and SDF- 1αKDEL have no significant deviation for phenotypic knockout of CXCR4. These suggest that the phenotypic knockout effects of SDF-1α/54 against CXCR4 are not influenced by deleting of SDF-1α helix in the C-terminal.

关 键 词:细胞学 细胞诱导因子 SDF-1α/54 CXCR4
文章编号:1004-0552(2007)03-0111-06
修稿时间:2007-04-25
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