Construction of Sense and Antisense of Human Heparanase and Green Fluorescent Protein Eukaryotic Co-expressing Vectors and Their Expression in Pancreatic Cancer Cell Lines SW1990
- VernacularTitle:正、反义Heparanase基因荧光真核表达载体的构建及胰腺癌SW1990细胞转染
- Author:
Yan YANG
- Publication Type:Journal Article
- Keywords:
Heparanase;
Fluorescent eukaryotic expressing vector;
Transfection
- From:
Journal of Kunming Medical University
1990;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct sense and antisense of human heparanase and green fluorescent protein eukaryotic expressing vectors and then transfect these vectors into Pancreatic Cancer Cell Lines SW1990.Methods The human Heparanase cDNA fragment contained in the pcDNA3 Hpa vector was cloned into the enhanced green fluorescent protein eukaryotic expressing vector pIRES2-EGFP in cis-direction or trans-direction.The recombinant vectors were identified by digestion of BamH I and further identified by DNA sequencing.The recombinant were transfected to pancreatic cancer cells SW1990 by liposome method.After 24h,the transfected cells were observed under fluorescent inverted microscope.Results After digested by BamH I,two fragments with the length of 5.3 kb and 1.7 kb were formed in sense fluorescent eukaryotic expressing vector,while two other fragments with the length of 6.5 kb and 0.5 kb were formed in antisense fluorescent vector.The DNA sequencing was also confirmed the linking direction of sense and antisense recombinant.Green fluorescence of the transfected cells could be observed under fluorescent inverted microscope after 48h of transfection.Conclusion Human heparanase sense and antisense fluorescent eukaryotic expressing vectors are successfully constructed and transfected to human pancreatic cancer cell lines.
