Cloning of the Promoter Regions of Mouse TGF-{beta} Receptor Genes by Inverse PCR with Highly Overlapped Primers |
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Authors: | Yoshitomo-Nakagawa, Kiyomi Muramatsu, Masa-aki Sugano, Sumio |
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Affiliation: | Department of Virology, Institute of Medical Science, University of Tokyo 4-6-1, Shirokanedai, Minato-ku, Tokyo 108 1Helix Research Institute 1532-3 Yana, Kisarazu-shi, Chiba 292, Japan |
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Abstract: | In order to isolate promoters of mouse TGF-ß receptorgenes, we used inverse PCR with highly overlapped primers correspondingto the 5' sequence of the receptor cDNAs. Nested primer setsonly covered a 30- to 40-base region of the sequences. HinfI-digestedand self-ligated mouse genomic DNA was used as a PCR template.Only one band for each receptor was seen after PCR. The amplifiedDNA fragments could direct luciferase production when the luciferasecoding sequence was ligated after the fragments. The sequenceof the fragment which correspond to the type II receptor showedpartial homology with the promoter region of the human TGF-ßtype II receptor. Thus, the inverse PCR with highly overlappedprimers could be an easy way to isolate the promoter regionsof many genes. |
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Keywords: | inverse PCR TGF-ß receptor promoter cDNA 5'-end |
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