2011•PubMedRequires access
[Construction of eukaryotic expression vector of human CD34 and establishment of stably transfected cell line].
Lin Shi, Hao Qu, Hongqin Liu, Hai-xia Gao, Weibin Zhu
Open publisher page 1 citations
Abstract
AIM: To construct eukaryotic expression vector of human CD34 and transfect it to 3T3 cells so as to establish stably transfected 3T3 cell line. METHODS: The RNA was extracted from KG1a. CD34 gene was amplified by RT-PCR. With the double-enzyme digestion, CD34 gene was cloned into pCI-neo eukaryotic expression vector, yielding pCI-CD34. The pCI-CD34 was transfected into 3T3 cell by electroporator. Stably transfected 3T3 cell line was established, and the CD34 expression in the transfected cells was detected by RT-PCR and FACS. RESULTS: The eukaryotic expression vector pCI-CD34 was constructed, and stably transfected 3T3 cell line was established. CONCLUSION: Construction of eukaryotic expression vector of CD34 and the establishment of stably transfected 3T3 cell line are helpful to preparation of anti-CD34 mAbs and further functional study of CD34.