Cloning, subcellular localization and in situ detection of Xenopus laevis beta-synnclein gene]

To clone Xenopus laevis beta-synuclein gene (xSYNB) and study the subcellular localization of xSYNB protein.According to the xSYNB cDNA sequence published in GenBank, a pair of primers were designed. The encoding region of xSYNB gene from the adult Xenopus laevis brain was amplified by RT-PCR, and then was cloned into pGEM-T Easy vector. The resulting recombinant plasmid was named as TA-xSYNB. The xSYNB cDNA was further subcloned into the pEGFP-N1 vector, and the resulting recombinant expression plasmid pEGFPN1-xSYNB was transfected into HEK293 cells to analyze the subcellular localization of xSYNB. The whole-mount in situ hybridization of embryos were used to identify the expression localization of xSYNB.The recombinant expression plasmid pEGFPN1-xSYNB was successfully constructed. The results of green fluorescence detection suggested that xSYNB gene was mainly expressed in the cytoplasm, and the results of in situ hybridization suggested that xSYNB gene was mainly expressed in the brain of embryo.The xSYNB gene may play a role in the development of the nervous system of Xenopus laevis.