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/10 6 cell/24 h (GlNaCcIX) and 211 ng/10 6 cell/24 h (GlNaMBcIX) respectively. Those data offered a promising result for further animal study.

Original languageEnglish (US)
Pages (from-to)374-375
Number of pages2
JournalScience in China, Series C: Life Sciences
Volume42
Issue number4
StatePublished - Aug 1 1999
Externally publishedYes

Fingerprint

retroviral vectors
blood coagulation factors
Factor IX
Blood Coagulation Factors
Cloning
Canidae
Organism Cloning
molecular cloning
skin
Complementary DNA
Fibroblasts
dogs
Skin
plasmid
Oligonucleotides
Liver
Actins
animal
Animals
Plasmids

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)

Cite this

Cloning and expression of canine clotting factor IX cDNA in vitro mediated by retroviral vector. / Gao, Xiaobo; Qiu, Xinfang; Lu, Daru; Xue, Jinglun.

In: Science in China, Series C: Life Sciences, Vol. 42, No. 4, 01.08.1999, p. 374-375.

Research output: Contribution to journalArticle

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AU - Gao, Xiaobo

AU - Qiu, Xinfang

AU - Lu, Daru

AU - Xue, Jinglun

PY - 1999/8/1

Y1 - 1999/8/1

N2 - 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/10 6 cell/24 h (GlNaCcIX) and 211 ng/10 6 cell/24 h (GlNaMBcIX) respectively. Those data offered a promising result for further animal study.

AB - 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/10 6 cell/24 h (GlNaCcIX) and 211 ng/10 6 cell/24 h (GlNaMBcIX) respectively. Those data offered a promising result for further animal study.

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