Construction of two recombination yeast two-hybrid vectors by in vitro recombination
| العنوان: | Construction of two recombination yeast two-hybrid vectors by in vitro recombination |
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| المؤلفون: | Yingtong Wang, Feng Guo, Yuzhu Zhang |
| المصدر: | Molecular Biotechnology. 36:38-43 |
| بيانات النشر: | Springer Science and Business Media LLC, 2007. |
| سنة النشر: | 2007 |
| مصطلحات موضوعية: | Recombination, Genetic, Cloning, Genetics, Base Sequence, cDNA library, Genetic Vectors, Molecular Sequence Data, Bioengineering, Saccharomyces cerevisiae, Biology, Applied Microbiology and Biotechnology, Biochemistry, WBP11, Two-Hybrid System Techniques, Restriction digest, Vector (molecular biology), Site-specific recombination, Molecular Biology, In vitro recombination, Recombination, Plasmids, Biotechnology |
| الوصف: | Recombination-based restrictionless, ligation-independent cloning has been proven to be advantageous over restriction digestion and ligation cloning. To utilize the recombination cloning and previously constructed two-hybrid cDNA libraries, a new Gateway yeast two-hybrid bait vector, pEZY202, and a new prey vector, pEZY45, were constructed. The two-hybrid vectors were generated by in vitro recombination using a protocol that can be easily adapted for the conversion of other existing vectors. The new vectors were used to assay the interaction between the WW domain of PQBP1 (PQBPww) and the WW domain binding protein WBP11. Both PQBPww and WBP11 were cloned into a Gateway donor vector by in vitro recombination. They were then subcloned into pEZY45 and pEZY202, respectively, by in vitro recombination. The binding between PQBPww and WBP11 was reported in a two-hybrid experiment using the new vectors. The results of testing the new vectors in combination with the original vectors indicated that the new bait vector could be used to screen cDNA libraries that are constructed using the original prey vectors. |
| تدمد: | 1559-0305 1073-6085 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1e2991e39160a9e82257ed14da2e9211 https://doi.org/10.1007/s12033-007-0018-0 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....1e2991e39160a9e82257ed14da2e9211 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 15590305 10736085 |
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