Correction of RNA aberrant splice increases foreign gene expression in transgenic mice |
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Authors: | Yifan Lu Chai Tian Jixian Deng Peitang Huang |
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Institution: | (1) Institute of Biotechnique, Academy of Military Medical Science, 100071 Beijing, China;(2) Cardiovascular Institute and Fuwai Hospital, Chinese Academy of Medical Sciences, 100037 Beijing, China |
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Abstract: | Transgenic mice with mammary gland secreting human granulocyte colony stimulating factor (G-CSF) were produced using mice
whey acid protein gene promoter. It was found that there was very low expression level in mammary gland. Human G-CSF cDNA
was obtained by RT-PCR from transgenic mice mammary gland. Sequence analysis showed that this G-CSF gene deleted the 4th exon,
and compared with human G-CSF genomic DNA, there were donor and acceptor splice sites in the deletion fragment. It was considered
that the 3rd and 4th introns also delete in G-CSF fragment. The transgenic construct was corrected by deleting the 3rd and
4th introns to construct the minigene, which was used to produce transgenic mice by microinjection. Northern blot showed that
G-CSF expression using the new construct increased 5.4 times as that before in transgenic mice. The results suggested that
it was possible that RNA aberrant splice result in low expression in transgenic mice. |
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Keywords: | transgenic RNA aberrant splice expression |
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