Construction of pEGFP-C1/U6-mediated plasmid expressing MDR1 shRNA.
- Author:
Xi-Bin XIAO
1
;
Zhao-Xia XIE
Author Information
1. Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
ATP-Binding Cassette, Sub-Family B, Member 1;
biosynthesis;
genetics;
DNA-Binding Proteins;
Drug Resistance, Multiple;
genetics;
Drug Resistance, Neoplasm;
genetics;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
genetics;
Humans;
Plasmids;
genetics;
RNA, Messenger;
biosynthesis;
genetics;
RNA, Small Interfering;
biosynthesis;
genetics;
RNA, Small Nuclear;
genetics
- From:
Journal of Experimental Hematology
2006;14(2):384-387
- CountryChina
- Language:Chinese
-
Abstract:
To construct a plasmid expressing MDR1 short hairpin RNA (shRNA) mediated by pEGFP-C1/U6 vector, two coding sequences of 19 nucleotides were selected from MDR. Two pairs of oligonucleotides were designed for these two fragments. After annealing the formed double-stranded DNAs were ligated with plasmid pEGFP-C1/U6 (pEGFP-C1 vector with U6 promoter). The plasmids producing MDR1 shRNA were constructed from the inverted motif containing 9 spacers and four Ts. The results showed that the constructed plasmids were named pEGFP-C1/U6/A and pEGFP-C1/U6/B, and the constructs were identified by restriction and sequence analysis, no any base mutation was observed. It is concluded that plasmids of pEGFP-C1/U6/A and pEGFP-C1/U6/B expressing MDR1 shRNA were successfully constructed, providing a highly effective method for reversing the multidrug resistance in clinic.
