Cloning and expression of canine clotting factor IX cDNA in vitro mediated by retroviral vector

Xiaobo Gao, Xinfang Qiu, Daru Lu, Jinglun Xue

Research output: Contribution to journalArticle

Abstract

Oligonucleotide of cFIX cDNA (canine FIX, cFIX) was used to transcript mRNA of dog liver cell to cDNA by RT-PCR, and further construct it on the plasmid vector pGEM-T. The correct sequence of cFIX cDNA was obtained which covered the entire cFIX coding region. Furthermore, GlNaCcIX (driven by hCMV promoter) and GlNaMBcIX (driven by MCK enhancer and β-actin promoter) were constructed using the retroviral vector backbone of GlNa. Canine skin fibroblast (CSF) was used as target cell, transduced with the above constructors respectively. The results showed that these modified CSF cells could express cFIX and that the expression levels were 173 ng/106 cell/24 h (GlNaCcIX) and 211 ng/106 cell/24 h (GlNaMBcIX) respectively. Those data offered a promising result for further animal study.

Original languageEnglish (US)
Pages (from-to)370-375
Number of pages6
JournalScience in China, Series C: Life Sciences
Volume42
Issue number4
DOIs
StatePublished - Aug 1999

Keywords

  • Canine clotting factor IX
  • Canine skin fibroblast
  • Gene expression
  • Gene transfer
  • Retroviral vector

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)
  • Agricultural and Biological Sciences(all)

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