Construction of lentivirus carrying novel human pancreatic cancer gene S100P and green fluorescent protein gene
10.3724/SP.J.1008.2008.01230
- Author:
Gang LI
1
Author Information
1. Department of General Surgery
- Publication Type:Journal Article
- Keywords:
Green fluorescent protein;
Lentivirus;
Pancreatic neoplasms;
S100P gene
- From:
Academic Journal of Second Military Medical University
2010;29(10):1230-1233
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct lentivirus carrying both novel human pancreatic cancer gene S100P and green fluorescent protein (GFP) gene. Methods: The fragments containing all the exons of S100P were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP. The lentivirus was packaged and was used to transfect 293T cells together with pShuttle. The supernatant of virus-producing cells was harvested, concentrated, identified, and was used to infect 293T cells and pancreatic cancer cells. Fluorescent microscopy was used to observe the fluorescence in the 293T cells; and real time-PCR was used to examine the relative contents of S100P in pancreatic cancer cells. Results: Electrophoresis showed that the sequence of the RT-PCR product was consistent with the data of NCBI by DNA sequencing analysis. The lentivirus effectively transfected 293T cells. Strong green fluorescence was observed by fluorescent microscopy. The supernatant of lentivirus-transfected 293T cells effectively infected 293T cells and the relative content of S100P in the transfected pancreatic cancer cells was higher than that of control group. Conclusion: The lentivirus vector containing S100P-GFP recombinant gene have been successfully constructed, which provides a basis for further study of S100P function.
