XJIPC OpenIR  > 资源化学研究室
Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method
Ma, Hairong; Lu, Zili; Sun, Yi; Peng, Tao; Shuai, Zhiqiang; Ma, Yufang; Zhang, Yan; Wang, Liang; Cao, Xu; Wang, Hanqing
2007
Source PublicationJOURNAL OF REPRODUCTION AND DEVELOPMENT
ISSN0916-8818
Volume53Issue:1Pages:95-104
AbstractIn this study, we introduced a co-transfection method for the selection of donor nuclei in somatic cell-mediated nuclear transfer. Two vectors were constructed in our experiment. One was pMSCV-GFP carrying the neomycin-resistant gene (Neo(r)) and the green fluorescent protein (GFP) reporter gene; the other was pBC1-GFP carrying the mammary gland-specific promoter and target gene GFP. Ovine adult fibroblasts were co-transfected with pMSCV-GFP and pBC1-GFP. The data from this work demonstrated that the GFP genes in both vectors could successfully co-integrate into the genomes of ovine adult fibroblasts in three of the four transgenic cell clones assayed. Furthermore, PCR analysis of transgenic embryos proved that the GFP genes in both vectors could cointegrate into the genomes of the reconstructed embryos. Subsequently, analysis of the developmental rate of the reconstructed embryos after nuclear transfer indicated that the blastocyst rate from the co-transfected donor cells was similar (approximate 8 percent) to that from individual pMSCV-GFP transfected donor cells. The influence of co-transfection resulting in modification of donor nuclei on development of reconstructed embryos was also investigated. The results of flow cytometric analysis indicated that the co-transfected ovine fibroblasts had similar quiescent characteristics in terms of cell cycle (G0+G1 percent: 73.20 +/- 4.04) to the individual pMSCV-GFP transfected fibroblasts (G0+G1 percent: 70.77 +/- 1.19) after they were treated with serum starvation for five days. Our results suggest that the co-transfection method can be used for selection of donor cell clones in somatic cell-mediated gene transfer experiments. It can be potentially extended to applications related to expression of functional protein in mammary glands and other transgenic research relevant to nuclear transfer.
KeywordCell Cycle Co-transfection Mammary Gland-specific Promoter Ovine Adult Fibroblast Somatic Cell-mediated Nuclear Transfer
DOI10.1262/jrd.18082
Indexed BySCI
WOS IDWOS:000244465700011
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.xjipc.cas.cn/handle/365002/4122
Collection资源化学研究室
AffiliationChinese Acad Sci, Xinjiang Tech Inst Phys & Chem, Urumqi 830011, Peoples R China;Chinese Acad Sci, Lanzhou Inst Chem Phys, Lanzhou 730000, Peoples R China;Xinjiang Golden Cattle Biol Incorp, Urumqi 830011, Peoples R China;Xinjiang Med Univ, Urumqi 830011, Peoples R China;Univ Alabama, Dept Pathol, Birmingham, AL 35294 USA;China Agr Univ, State Key Labs Agrobiotechnol, Coll Biol Sci, Beijing 475001, Peoples R China
Recommended Citation
GB/T 7714
Ma, Hairong,Lu, Zili,Sun, Yi,et al. Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method[J]. JOURNAL OF REPRODUCTION AND DEVELOPMENT,2007,53(1):95-104.
APA Ma, Hairong.,Lu, Zili.,Sun, Yi.,Peng, Tao.,Shuai, Zhiqiang.,...&Wang, Hanqing.(2007).Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method.JOURNAL OF REPRODUCTION AND DEVELOPMENT,53(1),95-104.
MLA Ma, Hairong,et al."Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method".JOURNAL OF REPRODUCTION AND DEVELOPMENT 53.1(2007):95-104.
Files in This Item:
File Name/Size DocType Version Access License
Selection of donor n(820KB)期刊论文作者接受稿开放获取CC BY-NC-SAView Application Full Text
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Ma, Hairong]'s Articles
[Lu, Zili]'s Articles
[Sun, Yi]'s Articles
Baidu academic
Similar articles in Baidu academic
[Ma, Hairong]'s Articles
[Lu, Zili]'s Articles
[Sun, Yi]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Ma, Hairong]'s Articles
[Lu, Zili]'s Articles
[Sun, Yi]'s Articles
Terms of Use
No data!
Social Bookmark/Share
File name: Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method.pdf
Format: Adobe PDF
This file does not support browsing at this time
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.