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Grafting of Beads into Developing Chicken Embryo Limbs to Identify Signal Transduction Pathways Affecting Gene Expression
Published on: January 17, 2016
[Construction of chicken embryo fibroblasts cDNA expression library]
Wei Liu1, Yu-long Gao, Hong-lei Gao
1Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. lw-790613@126.com
Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica
|August 4, 2007
Summary
Researchers constructed a high-quality chicken embryo fibroblast cDNA expression library using Gateway technology. This library aids in understanding infectious bursal disease virus tropism and identifying cellular receptors in chickens.
Area of Science:
- Molecular Biology
- Virology
- Immunology
Background:
- Chicken embryo fibroblast (CEF) is crucial for studying infectious bursal disease virus (IBDV).
- Understanding IBDV cell tropism and receptors requires a robust CEF cDNA expression library.
- Traditional cloning methods present technical limitations for library construction.
Purpose of the Study:
- To construct a high-quality CEF cDNA expression library using Gateway technology.
- To overcome limitations of traditional cloning methods for IBDV research.
- To facilitate the identification of IBDV receptors and study cellular tropism.
Main Methods:
- mRNA extraction from chicken embryonic fibroblast.
- cDNA synthesis using biotin-conjugated Oligo (dT) primer.
- Gateway technology for BP and LR recombination reactions for library construction.
Main Results:
- A cDNA entry library was constructed with a titer of 1 x 10^6 cfu/mL and an average insert size of 2243 bp.
- An expression library was generated with a titer of 5 x 10^5 cfu/mL and an average insert size of 2411 bp.
- The library exhibited high recombination rates and coverage.
Conclusions:
- The constructed CEF cDNA expression library is of high quality.
- This resource can aid in identifying IBDV receptors and understanding cellular tropism.
- The library provides valuable transcriptional data for studying CEF biological functions.
