Developmental regulation of 14-3-3 ε isoform in rat heart
Sharon Luk, Sai Ming Ngai, Stephen Kwok‐Wing Tsui, Kwok-keung Chan, Kwok‐Pui Fung, Cheuk-yu Lee, Mary Miu Yee Waye
Abstract
Sharon Luk, Sai Ming Ngai, Stephen Kwok‐Wing Tsui, Kwok-keung Chan, Kwok‐Pui Fung, Cheuk-yu Lee, Mary Miu Yee Waye
Abstract
Human heart cDNA sequencing yielded a cDNA clone that is similar in DNA and amino acid sequences to that of mouse 14-3-3 epsilon isoform. The 6xHis-tagged H1433 epsilon recombinant protein was expressed in Escherichia coli and its size was approximately 30 kDa. From Northern blot results with human multiple tissues, human skeletal muscle was found to have the highest level of h1433 epsilon mRNA expression, whereas Northern blots of human cancer cell lines detected the highest mRNA level of h1433 epsilon in colorectal adenocarcinoma SW480. The protein expression level of h1433 epsilon and Raf-1 is found to be regulated coordinately during rat heart development, and their protein expression was highest from 14.5 to 16.5 days postcoitum.
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Human heart cDNA sequencing yielded a cDNA clone that is similar in DNA and amino acid sequences to that of mouse 14-3-3 epsilon isoform. The 6xHis-tagged H1433 epsilon recombinant protein was expressed in Escherichia coli and its size was approximately 30 kDa. From Northern blot results with human multiple tissues, human skeletal muscle was found to have the highest level of h1433 epsilon mRNA expression, whereas Northern blots of human cancer cell lines detected the highest mRNA level of h1433 epsilon in colorectal adenocarcinoma SW480. The protein expression level of h1433 epsilon and Raf-1 is found to be regulated coordinately during rat heart development, and their protein expression was highest from 14.5 to 16.5 days postcoitum.
Key concepts: Complementary DNA, Northern blot, Gene isoform, Molecular biology, Biology, Recombinant DNA, Messenger RNA, cDNA library