Construction of SMYD3 liver-specific expression transgene for the eestablishment of transgenic mice

摘要

SET and MYND domain-containing protein 3 (SMYD3) is a novel histone methyltransferase involved in human carcinogenesis. To further establish the transgenic mice of SMYD3 and provide a useful platform for the study of SMYD3 and the screening of novel antitumor drugs, a transgene for liver-specific simultaneous expression of SMYD3 and EGFP was constructed. The human SMYD3 gene was subcloned into pIRES2-EGFP eukaryotic expressing vector. Subsequently, mice albumin promoter (pALB) (−821∼+59) was obtained from the mice genome DNA by PCR and then inserted into the 5'' flanking region of SMYD3. The final recombinant plasmid (named as pIRAS) was digested by AflII StuI and XhoI , and the DNA fragment consisting of albumin promoter (pALB) SMYD3 Internal Ribosome Entry Site (IRES) Enhanced Green Fluorescent Protein (EGFP) and SV40 polyA signal sequence (named as ASIES) was released. The linearized, purified ASIES transgene was microinjected into C57BL/6 mouse zygotes. After cultured for 2 days in vitro, most of the injected zygotes showed normal division, and the following works would establish the transgenic mice.