Cloning of full-length cDNA of teleost corticotropin-releasing hormone precursor by improved inverse PCR.
We designed a new inverse PCR protocol combined with switching mechanism at 5' end of RNA transcript (SMART) technology, and applied it to the cloning of teleost corticotropin-releasing hormone precursor cDNA. Due to the advantages of both techniques, this method can efficiently amplify the complete 5'- and 3'-ends of cDNA in a single reaction, and might prove to be an alternative to the conventional rapid amplification of cDNA ends (RACE) approaches.
Kuniyoshi
Hisato
H
Graduate School of Biosphere Science, Hiroshima University, Japan. hkuni@hiroshima-u.ac.jp
Fukui
Yukio
Y
Sakai
Yoichi
Y
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003346:Corticotropin-Releasing Hormone / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics
D018076:DNA, Complementary; D011498:Protein Precursors; D012333:RNA, Messenger; D003346:Corticotropin-Releasing Hormone