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Objective: This paper studies the characteristics of the new human gene LACE1 and function of the gene cloning, expression and functional analysis, this study has both theoretical significance but also has potential applications for the diagnosis and treatment of disease provide a theoretical basis more will research and develop our own intellectual property means of gene therapy and genetic engineering drugs lay the theoretical foundation and provide new ideas. Methods: Analysis of bioinformatics software LACE1 gene sequence, gene expression and species of LACE1 homology analysis and prediction intracellular localization; detected by RT-PCR method LACE1 gene expression in different cells and to establish the target gene eukaryotic in vitro transfection, expression platform; using molecular cloning and expression of the target gene LACE1 constructed plasmid; using confocal microscopy further testing LACE1 gene subcellular localization; by morphological observation, MTT and other means to study gene overexpression LACE1 293T cell proliferation implications, using western blot detection after overexpression LACE1 and apoptosis related proteins caspase impact. Results: 1 through bioinformatics analysis LACE1 (Homo sapiens lactation elevated 1), which GeneID: 246269; RefSeq: NM 1 45315.3. The gene is 2262 bp, consists of 13 exons and 12 introns, positioning and 6q 21. from 187 ~ 1632 encoding 481 amino acids with an open reading frame encoding a protein of about 54 kDa. The protein isoelectric point of 7.29, no disulfide bonds. 2 bioinformatics analysis software Biogps expression profiles predict and RT-PCR showed LACE1 is ubiquitously expressed, including in Hela cells, HepG2 cells with high expression, Raji cells, 293 cells, H520 cells showed moderate expression, 293T cells with low expression, suggesting that the gene may be associated with reduced cancer-related signaling pathways. 3 BLASTn in NCBI database through comparative analysis, which found LACE1 in human (Homo sapiens), cattle (Bos taurus), dog (Canis lupus fami), mouse (Mus musculis), rat (Rattus norvegicu), chicken ( Gallus gallus) and many other species homology exists. 4 would LACE1 gene amplified by PCR and cloned into the eukaryotic expression vector pcDNA3.1B (pcDB), by restriction enzyme digestion and sequencing proved that gene sequences in GeneBank sequence consistent with the published database that pcDB-LACE1 expression plasmid was constructed successfully. And constructed a plasmid carrying green fluorescent labels pEGFP-N1-LACE1. 5. Subcellular localization showed LACE1 gene was localized in the cytoplasm. 6 overexpression LACE1, 293T cells showed morphological death off phenomenon, MTT test results showed that the gene can cause cell death, western blot showed that the gene induced cell death may be caspase-dependent. Conclusions: 1. Discovered a novel human genes evolutionarily conserved LACE1, its full-length cDNA of 2262bp, with 13 exons and 12 introns, mainly located in the human chromosome 6, the gene is located in the cytoplasm. (2) the gene in normal and abnormal human tissues were expressed, indicating widespread expression of the gene, discovered by RT-PCR in Hela cells, HepG2 cells highly expressed in 293 cells, 293T cells showed low expression status, suggesting that Gene expression in tumor tissue than in normal tissue. 3. LACE1 can promote apoptosis, caspase-dependent and may be.
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