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Journal Article

Journal Article

Preparation and amplification of colony of goat transgenic fetal fibroblast and mammary gland epithelial cell with human lactoferrin gene  [2009]

Zhang Yuling, Henan University of Science and Technology, Luoyang (China), College of Animal Science and Technology; Liu Fengjun, Henan University of Science and Technology, Luoyang (China), College of Animal Science and Technology; Zhang Yong, Northwest Agricultural and Foresty University, Yangling (China), Bioengieering Institute;

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[Objective] The aim was to explore technical system of making single transgenic positive cells become colony cells by amplification culture. [Method] Fetal fibroblasts and mammary gland epithelial cells of single goat fetus of pBLM-C1 which specifically expressed human lactoferrin were cloned. Single cell colony of single transfection cell was prepared with 3 concentrations of 0%, 50% and 100% conditioned culture media. Transfection cell and non-transfection cell were carried out amplification culture by con-culture, neo gene was as screened gene, genome DNA of transfection cell was detected by PCR method. Chromosome karyotype analysis of single colony cell was tested. [Result] Compared with non-conditioned culture medium, 100% conditioned culture medium could greatly increase survived rate of single colony cells (FF: 53.33% vs. 10.00%; MGE: 33.33% vs. 6.67%). Compared with control, con-culture of transfection cell and non-transfection cell could greatly increase rate of transfection cell single colony after amplification culture (FF: 53.33% vs. 10.00%;MGE: 33.33% vs. 6.67%), confluence time of amplification culture was significantly decreased (20-30 d). The result of PCR showed that the colony cell obtained by above method contained hLF target gene. The result of karyotype analysis showed that most cloned cell chromosomes were normal. [Conclusion] The study provides a reliable method for separating transgenic cell, inserting and diagnosing ideal vector, a
nd can save expense and time for transgenic animal production.
From the journal
Agricultural Science and Technology
ISSN : 1009-4229

Bibliographic information

Language:
English
Type:
Journal Article
In AGRIS since:
2010
Volume:
10
Issue:
4
Start Page:
27
End Page:
30 100
All titles:
"Preparation and amplification of colony of goat transgenic fetal fibroblast and mammary gland epithelial cell with human lactoferrin gene"@eng
"制备和转基因山羊胎儿成纤维细胞和乳腺上皮细胞集落与人乳铁蛋白基因的扩增"@zho
Other:
"Resume(En)"
"13 ref."
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Bibliographic information

Language:
English
Type:
Journal Article
In AGRIS since:
2010
Volume:
10
Issue:
4
Start Page:
27
End Page:
30 100
All titles:
"Preparation and amplification of colony of goat transgenic fetal fibroblast and mammary gland epithelial cell with human lactoferrin gene"@eng
"制备和转基因山羊胎儿成纤维细胞和乳腺上皮细胞集落与人乳铁蛋白基因的扩增"@zho
Other:
"Resume(En)"
"13 ref."