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Construction of eukaryotic expression vector of thrombospondin-1 type I repeat sequence
Xiao-Wei Li1, Yi-Li Liu, Ping-Sheng Wu
1Department of Cardiovascular Diseases, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Objective:
To construct eukaryotic expression vector of thrombospondin-1 type I repeat sequences.
Methods:
Thrombospondin-1 type I repeat sequence gene was amplified from human fetal lung tissue by reverse transcriptase-PCR (RT-PCR) to construct both recombinant clone vector and expression vector through coupling reaction, followed by transforming these vectors into E.coli DH5alpha. The positive clones were selected for verification by double enzyme digestion and sequence analysis.
Result:
The expected amplification product, thrombospondin-1 type I repeat gene sequence, was acquired by RT-PCR, which had a consistency up to 99% with the sequence from the GenBank.
Conclusion:
The eukaryotic expression vector PcDNA3.1(+)/TSP-1 type I repeat sequence was successfully constructed.
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