[Construction and expression of recombinant eucaryotic expression plasmid of amastin gene of Leishmania Donovani].
OBJECTIVE
To construct recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani and detect expression of the gene in NIH3T3 cells.


METHODS
Amastin gene was amplified from nuclear DNA of Leishmania Donovani isolates and cloned into an eukaryotic expression vector pcDNA3.1(+). The recombinant plasmid was named pcDNA3.1-amastin. NIH3T3 cell was transfected by pcDNA3.1-amastin. Transient and stable expression of amastin gene were detected by immunofluoresence and RT-PCR.


RESULTS
It was found that there was high green fluorescence on the cell membrane and inside the cell. It showed that NIH3T3 cell was transfected by pcDNA3.1-amastin successfully.


CONCLUSIONS
A recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani was successfully constructed, and can be expressed stably in the NIH3T3 cells.
Li
Jin-Fu
JF
Department of Parasitology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China. lijinfu72@163.com
Chen
Jian-Ping
JP
Yang
Zhi-Wei
ZW
Tian
Yu
Y
Ma
Ying
Y
Hu
Xiao-Su
XS
D000818:Animals; D005455:Fluorescent Antibody Technique; D007893:Leishmania donovani / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins