Construction of recombinant eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes
10.3980/j.issn.1672-5123.2014.12.11
- VernacularTitle:脂质体包裹人促甲状腺激素受体胞外段基因真核表达质粒的构建
- Author:
Yu-Li, YANG
;
Qing-Li, LUO
;
Hong-Bin, LV
- Publication Type:Journal Article
- Keywords:
human thyrotropin receptor extracellular domain gene;
shuttle vector PHMCMVTSHR289;
eukaryotic expression plasmid pcDNA3. 1 +;
recombination plasmid pcDNA3. 1+/TSHR289;
cationic liposomes
- From:
International Eye Science
2014;(12):2151-2154
- CountryChina
- Language:Chinese
-
Abstract:
AlM:To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes.
METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3. 1 +, and accredited whether pcDNA3. 1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3. 1+/TSHR289.
RESULTS: Recombination plasmid pcDNA3. 1+/TSHR289 digested with enzyme Hindlll and the fragment through 0. 8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3. 1+/TSHR289 were found synonymous mutation through forward ( AAC to AAT ) and reverse sequencing ( GCG to GCT) . The volume ratio of cationic liposomes and recombinant plasmid was 3:1.
CONCLUSlON: lt is successful to construct the recombination plasmid pcDNA3. 1+/TSHR289 by accredit it through enzymatic digestion and sequencing.
