Efficient production of transgenic goat (Capra hircus) embryos using dual markers

Liang Wang; Tingting Liu; Tao Peng; Dajiang Zhang; Hanqing Wang; Xu Cao; Weihua Li; Anjiang Wang; Hai Zhu

doi:10.1016/j.smallrumres.2007.05.007

Efficient production of transgenic goat (Capra hircus) embryos using dual markers

Liang Wang, Tingting Liu, Tao Peng, Dajiang Zhang, Hanqing Wang, Xu Cao, Weihua Li, Anjiang Wang, Hai Zhu

Research output: Contribution to journal › Article › peer-review

Abstract

The objective of this study was to develop an effective method to screen transgenic donor cells and improve the production efficiency of transgenic embryos. In the study, goat (Capra ibex) skin fibroblasts were co-transfected with two expression constructs, one containing the human α1 (I) procollagen cDNA fragment (4.3 kb) and the neomycin resistance (Neo^r) gene, and the second containing the green fluorescent protein (GFP) gene. The Neo^r and GFP genes were used as markers for the screening of donor cells. Results revealed a significant difference (P < 0.01) in the production rate of positive transgenic blastocysts (44.6% versus 82.8%) between donor fibroblasts selected using antibiotic G418 alone (Treatment A) and those from donor fibroblasts selected using antibiotic G418 and GFP (Treatment B), respectively. No significant differences were found between Treatments A and B in the total somatic cell nuclear transfer (SCNT) embryo cleavage rates (71.1% versus 65.9%) and total SCNT blastocyst formation rates (18.2% versus 15.7%). The research results indicate the Neo^r and GFP genes located at different constructs can be effectively combined by co-transfection to screen transgenic donor fibroblasts, and increase the yield of transgenic SCNT embryos carrying human α1 (I) procollagen cDNA in goats.

Original language	English (US)
Pages (from-to)	99-104
Number of pages	6
Journal	Small Ruminant Research
Volume	75
Issue number	1
DOIs	https://doi.org/10.1016/j.smallrumres.2007.05.007
State	Published - Feb 2008
Externally published	Yes

Keywords

Embryos
Fibroblasts
Goats
Green fluorescent protein
Neomycin resistance gene
Nuclear transfer

ASJC Scopus subject areas

Food Animals
Animal Science and Zoology

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10.1016/j.smallrumres.2007.05.007

Cite this

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title = "Efficient production of transgenic goat (Capra hircus) embryos using dual markers",

abstract = "The objective of this study was to develop an effective method to screen transgenic donor cells and improve the production efficiency of transgenic embryos. In the study, goat (Capra ibex) skin fibroblasts were co-transfected with two expression constructs, one containing the human α1 (I) procollagen cDNA fragment (4.3 kb) and the neomycin resistance (Neor) gene, and the second containing the green fluorescent protein (GFP) gene. The Neor and GFP genes were used as markers for the screening of donor cells. Results revealed a significant difference (P < 0.01) in the production rate of positive transgenic blastocysts (44.6% versus 82.8%) between donor fibroblasts selected using antibiotic G418 alone (Treatment A) and those from donor fibroblasts selected using antibiotic G418 and GFP (Treatment B), respectively. No significant differences were found between Treatments A and B in the total somatic cell nuclear transfer (SCNT) embryo cleavage rates (71.1% versus 65.9%) and total SCNT blastocyst formation rates (18.2% versus 15.7%). The research results indicate the Neor and GFP genes located at different constructs can be effectively combined by co-transfection to screen transgenic donor fibroblasts, and increase the yield of transgenic SCNT embryos carrying human α1 (I) procollagen cDNA in goats.",

keywords = "Embryos, Fibroblasts, Goats, Green fluorescent protein, Neomycin resistance gene, Nuclear transfer",

author = "Liang Wang and Tingting Liu and Tao Peng and Dajiang Zhang and Hanqing Wang and Xu Cao and Weihua Li and Anjiang Wang and Hai Zhu",

note = "Funding Information: This study was financially supported by the National Natural Science Foundation of China (Grant No. 30471242). ",

year = "2008",

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doi = "10.1016/j.smallrumres.2007.05.007",

language = "English (US)",

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pages = "99--104",

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T1 - Efficient production of transgenic goat (Capra hircus) embryos using dual markers

AU - Wang, Liang

AU - Liu, Tingting

AU - Peng, Tao

AU - Zhang, Dajiang

AU - Wang, Hanqing

AU - Cao, Xu

AU - Li, Weihua

AU - Wang, Anjiang

AU - Zhu, Hai

N1 - Funding Information: This study was financially supported by the National Natural Science Foundation of China (Grant No. 30471242).

PY - 2008/2

Y1 - 2008/2

N2 - The objective of this study was to develop an effective method to screen transgenic donor cells and improve the production efficiency of transgenic embryos. In the study, goat (Capra ibex) skin fibroblasts were co-transfected with two expression constructs, one containing the human α1 (I) procollagen cDNA fragment (4.3 kb) and the neomycin resistance (Neor) gene, and the second containing the green fluorescent protein (GFP) gene. The Neor and GFP genes were used as markers for the screening of donor cells. Results revealed a significant difference (P < 0.01) in the production rate of positive transgenic blastocysts (44.6% versus 82.8%) between donor fibroblasts selected using antibiotic G418 alone (Treatment A) and those from donor fibroblasts selected using antibiotic G418 and GFP (Treatment B), respectively. No significant differences were found between Treatments A and B in the total somatic cell nuclear transfer (SCNT) embryo cleavage rates (71.1% versus 65.9%) and total SCNT blastocyst formation rates (18.2% versus 15.7%). The research results indicate the Neor and GFP genes located at different constructs can be effectively combined by co-transfection to screen transgenic donor fibroblasts, and increase the yield of transgenic SCNT embryos carrying human α1 (I) procollagen cDNA in goats.

AB - The objective of this study was to develop an effective method to screen transgenic donor cells and improve the production efficiency of transgenic embryos. In the study, goat (Capra ibex) skin fibroblasts were co-transfected with two expression constructs, one containing the human α1 (I) procollagen cDNA fragment (4.3 kb) and the neomycin resistance (Neor) gene, and the second containing the green fluorescent protein (GFP) gene. The Neor and GFP genes were used as markers for the screening of donor cells. Results revealed a significant difference (P < 0.01) in the production rate of positive transgenic blastocysts (44.6% versus 82.8%) between donor fibroblasts selected using antibiotic G418 alone (Treatment A) and those from donor fibroblasts selected using antibiotic G418 and GFP (Treatment B), respectively. No significant differences were found between Treatments A and B in the total somatic cell nuclear transfer (SCNT) embryo cleavage rates (71.1% versus 65.9%) and total SCNT blastocyst formation rates (18.2% versus 15.7%). The research results indicate the Neor and GFP genes located at different constructs can be effectively combined by co-transfection to screen transgenic donor fibroblasts, and increase the yield of transgenic SCNT embryos carrying human α1 (I) procollagen cDNA in goats.

KW - Embryos

KW - Fibroblasts

KW - Goats

KW - Green fluorescent protein

KW - Neomycin resistance gene

KW - Nuclear transfer

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Efficient production of transgenic goat (Capra hircus) embryos using dual markers

Abstract

Keywords

ASJC Scopus subject areas

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