[Construction of LACK gene recombinant plasmid and detection of its expression in eukaryotic cell].
The LACK gene from Leishmania, an analogue of the receptor of activated protein kinase C, was discovered recently. In this study, the LACK gene of Leishmania donovani was obtained from the recombinant plasmid T-LACK by PCR. The gene was cloned into eukaryotic expressed plasmid pcDNA3.1(+) to construct recombinant plasmid. This recombinant plasmid then was transfected into the eukaryotic cell COS-7, and the expression of LACK gene in eukaryotic cell was detected by RT-PCR and immunofluorescent staining. Both RT-PCR and immunofluorescent staining of recombinant plasmid transfected COS-7 showed positive reaction, thus indicating that the recombinant plasmid pcDNA3-LACK can express LACK protein in euka ryotic cell COS-7.
Ma
Ying
Y
Department of Parasitology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Hu
Xiaosu
X
Wang
Yajing
Y
Bu
Lingyi
L
D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D019556:COS Cells; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000235:genetics*; D005057:Eukaryotic Cells / Q000378:metabolism; D005822:Genetic Vectors; D007893:Leishmania donovani; D010957:Plasmids / Q000235:genetics*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D019444:Vaccines, DNA
D000953:Antigens, Protozoan; D004274:DNA, Recombinant; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D019444:Vaccines, DNA; C093139:LACK antigen, Leishmania