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Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
Published on: April 26, 2017
[Alternative splicing of SmU2AF65 in Solanum melongena L]
1Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan 430074, China. luoyan1984@126.com
Abstract:
U2AF (U2 snRNP auxiliary factor), an essential auxiliary factor for pre-mRNA splicing, is highly conserved across species. Here we report the cloning and expression analysis of the homologous gene (SmU2AF65) encoding the large subunit of U2AF65 in Solanum melongena. Gene specific primers for Rapid Amplification of cDNA Ends (RACE) are designed according to the genomic sequence of eggplant BAC 77N19 and the full length cDNA of tobacco NpU2AF65 a and b. The full length cDNA of SmU2AF65 (GenBank accession No. EU543263) contains a complete open reading fragment encoding a 554 amino acid protein, and the deduced protein contains three conserved RNA recognition domains (RRM). RT-PCR assays indicate that SmU2AF65 is constitutively expressed in leaf, flower, and root, with a different transcript found in root originated from alternative splicing.
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