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

Xiaobo Gao; Xinfang Qiu; Daru Lu; Jinglun Xue

doi:10.1007/bf02882056

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	370-375
Number of pages	6
Journal	Science in China, Series C: Life Sciences
Volume	42
Issue number	4
DOIs	https://doi.org/10.1007/bf02882056
State	Published - Aug 1999
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Environmental Science
General Agricultural and Biological Sciences

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10.1007/bf02882056

Cite this

@article{589b601c7dde4f0aaa0441bea6297da7,

title = "Cloning and expression of canine clotting factor IX cDNA in vitro mediated by retroviral vector",

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.",

keywords = "Canine clotting factor IX, Canine skin fibroblast, Gene expression, Gene transfer, Retroviral vector",

author = "Xiaobo Gao and Xinfang Qiu and Daru Lu and Jinglun Xue",

year = "1999",

month = aug,

doi = "10.1007/bf02882056",

language = "English (US)",

volume = "42",

pages = "370--375",

journal = "Science in China, Series C: Life Sciences",

issn = "1006-9305",

publisher = "Science in China Press",

number = "4",

}

TY - JOUR

T1 - Cloning and expression of canine clotting factor IX cDNA in vitro mediated by retroviral vector

AU - Gao, Xiaobo

AU - Qiu, Xinfang

AU - Lu, Daru

AU - Xue, Jinglun

PY - 1999/8

Y1 - 1999/8

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/106 cell/24 h (GlNaCcIX) and 211 ng/106 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/106 cell/24 h (GlNaCcIX) and 211 ng/106 cell/24 h (GlNaMBcIX) respectively. Those data offered a promising result for further animal study.

KW - Canine clotting factor IX

KW - Canine skin fibroblast

KW - Gene expression

KW - Gene transfer

KW - Retroviral vector

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U2 - 10.1007/bf02882056

DO - 10.1007/bf02882056

M3 - Article

C2 - 18763127

AN - SCOPUS:23044467660

SN - 1006-9305

VL - 42

SP - 370

EP - 375

JO - Science in China, Series C: Life Sciences

JF - Science in China, Series C: Life Sciences

IS - 4

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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