New pUC-derived expression vectors for rapid construction of cDNA libraries |
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| Authors: | I Oberb?umer |
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| Affiliation: | 1. Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-Ku, Shizuoka 422-8529, Japan;2. Department of Environmental Science, Faculty of Agriculture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh;1. Departamento de Zoologia e Botânica, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista, Cristóvão Colombo, 2265, 15054-000 São José do Rio Preto, SP, Brazil;2. Center for Brain, Behavior and Cognition, Department of Ecosystem Science and Management, The Pennsylvania State University, University Park, PA 16802, United States;3. Centro de Aquicultura da UNESP, Brazil;1. Department of Animal Science, University of Minnesota, St. Paul, MN;2. University of Minnesota Extension, Farmington, MN |
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| Abstract: | We have constructed a new series of the pUC-derived plasmids with an extended polylinker obtained from M13tg131 [Kieny et al., Gene 26 (1983) 91-99]. These vectors allowed us to design a simplified version of the method of Okayama and Berg [Mol. Cell. Biol. 2 (1982) 161-170] for establishing complete cDNA libraries. Improvements included easy recovery of the inserted cDNA due to the extended polylinkers; use of these vectors for gene expression in Escherichia coli, and amenability to supercoil sequencing with the universal primers of the M13 system [Chen and Seeburg, DNA 4 (1985) 165-170], which speeds up the identification of positive clones. Moreover, there is no need for an additional linker fragment, as was required by the Okayama and Berg [Mol. Cell. Biol. 2 (1982) 161-170] method. We have successfully used this system to obtain cDNA clones coding for the different chains of the large basement membrane proteins type IV collagen and laminin. |
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