Gene Cloning And Sequence Analysis Of Syntaxin 1B Of The Sea Perch

Exocytosis or endocrine is one of the most fundamental action of the organism. It maintains the life's normal metabolism and movement. It includes all kinds of the sides of the organism, such as growth, development and procreating. Exocytosis is a vital function of many cells including neuron, endocrine cell and immunocyte. Secretion in immunocytes involves a complex process of signal transduction. It is the fundament of life movement such as the release of neurotransmitter, secretion of hormone and transport of proteins. During the long time's evolution, the organism has developed a suit of faithful mechanism which ensures the life movemnet progresses work in order. This mechanism involoves every protein family of the SNARE complex.SNARE complex includes the core protein that is VAMP(synaptobrevin), SNAP25 and Syntaxin. So far, Syntaxin is the largest protein family in the SNARE complex. Syntaxin 1B is one of the Syntaxin isoforms. It has been cloned from many species of the mammals such as rat, mouse and human. In addition, it has been found from several marine biologies such as zebra fish, trout arid crab. But such research taking sea perch as the material has not been done in the world. This research is taking sea perch as the material, and cloning the Syntaxin 1B gene.After extracting RNA from sea perch tissue, we gained the first chain of cDNA by RT-PCR. Then designing a pair of primers according to the sequence of Syntaxin 1B from other organism. Based on the first chain of cDNA cloned from sea perch, the partial sequence of Syntaxin 1B gene is been cloned by using TD-PCR and we received the 751bp fragment at last. Then the fragment is eluted from the gel and is ligased with the pMD 18-T vector. It is transformed and cultured. Extracted plasmid is digested by Hind and Xbal. This result proves the ligation succeed. Blast the cloned sequence in the NCB net, we prove the sequence is the Syntaxin 1B gene. According to previous research of the Syntaxin 1B from other organism, the domain linking with BoNT/C is found from the Syntaxin 1B sequence. By the alignment, the gene's high homologous is found among many different biologies.So we deduce the Syntaxin 1B plays a important role in the process of vesicle transport in sea perch.