Transfer of an expression YAC into goat fetal fibroblasts by cell fusion for mammary gland bioreactor
Abstract
Yeast artificial chromosomes (YACs) as transgenes in transgenic animals are likely to ensure optimal expression levels. Microinjection of YACs is the exclusive technique used to produce YACs transgenic livestock so far. However, low efficiency and high cost are its critical restrictive factors. In this study, we presented a novel procedure to produce YACs transgenic livestock as mammary gland bioreactor. A targeting vector, containing the gene of interest-a human serum albumin minigene (intron 1, 2), yeast selectable marker (G418R), and mammalian cell resistance marker (neo{sup r}), replaced the {alpha}-lactalbumin gene in a 210 kb human {alpha}-lactalbumin YAC by homogeneous recombination in yeasts. The chimeric YAC was introduced into goat fetal fibroblasts using polyethylene glycol-mediated spheroplast fusion. PCR and Southern analysis showed that intact YAC was integrated in the genome of resistant cells. Perhaps, it may offer a cell-based route by nuclear transfer to produce YACs transgenic livestock.
- Authors:
-
- Shanghai Transgenic Research Center, No. 88 Cailun Road, Shanghai 201203 (China)
- Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433 (China)
- Publication Date:
- OSTI Identifier:
- 20710858
- Resource Type:
- Journal Article
- Journal Name:
- Biochemical and Biophysical Research Communications
- Additional Journal Information:
- Journal Volume: 333; Journal Issue: 1; Other Information: DOI: 10.1016/j.bbrc.2005.05.072; PII: S0006-291X(05)01057-0; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES; ALBUMINS; BLOOD SERUM; CHROMOSOMES; COST; EFFICIENCY; FIBROBLASTS; GENES; GOATS; MAMMARY GLANDS; POLYETHYLENE GLYCOLS; POLYMERASE CHAIN REACTION; TRANSGENIC ANIMALS; YEASTS
Citation Formats
Xufeng, Zhang, Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, Guoxiang, Wu, Chen, Jian-Quan, Aimin, Zhang, Siguo, Liu, Binghua, Jiao, and Guoxiang, Cheng. Transfer of an expression YAC into goat fetal fibroblasts by cell fusion for mammary gland bioreactor. United States: N. p., 2005.
Web. doi:10.1016/j.bbrc.2005.05.072.
Xufeng, Zhang, Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, Guoxiang, Wu, Chen, Jian-Quan, Aimin, Zhang, Siguo, Liu, Binghua, Jiao, & Guoxiang, Cheng. Transfer of an expression YAC into goat fetal fibroblasts by cell fusion for mammary gland bioreactor. United States. https://doi.org/10.1016/j.bbrc.2005.05.072
Xufeng, Zhang, Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, Guoxiang, Wu, Chen, Jian-Quan, Aimin, Zhang, Siguo, Liu, Binghua, Jiao, and Guoxiang, Cheng. 2005.
"Transfer of an expression YAC into goat fetal fibroblasts by cell fusion for mammary gland bioreactor". United States. https://doi.org/10.1016/j.bbrc.2005.05.072.
@article{osti_20710858,
title = {Transfer of an expression YAC into goat fetal fibroblasts by cell fusion for mammary gland bioreactor},
author = {Xufeng, Zhang and Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433 and Guoxiang, Wu and Chen, Jian-Quan and Aimin, Zhang and Siguo, Liu and Binghua, Jiao and Guoxiang, Cheng},
abstractNote = {Yeast artificial chromosomes (YACs) as transgenes in transgenic animals are likely to ensure optimal expression levels. Microinjection of YACs is the exclusive technique used to produce YACs transgenic livestock so far. However, low efficiency and high cost are its critical restrictive factors. In this study, we presented a novel procedure to produce YACs transgenic livestock as mammary gland bioreactor. A targeting vector, containing the gene of interest-a human serum albumin minigene (intron 1, 2), yeast selectable marker (G418R), and mammalian cell resistance marker (neo{sup r}), replaced the {alpha}-lactalbumin gene in a 210 kb human {alpha}-lactalbumin YAC by homogeneous recombination in yeasts. The chimeric YAC was introduced into goat fetal fibroblasts using polyethylene glycol-mediated spheroplast fusion. PCR and Southern analysis showed that intact YAC was integrated in the genome of resistant cells. Perhaps, it may offer a cell-based route by nuclear transfer to produce YACs transgenic livestock.},
doi = {10.1016/j.bbrc.2005.05.072},
url = {https://www.osti.gov/biblio/20710858},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 1,
volume = 333,
place = {United States},
year = {Fri Jul 22 00:00:00 EDT 2005},
month = {Fri Jul 22 00:00:00 EDT 2005}
}