This paper aims to provide better guidance for construction of trehalose-producing recombinant strains to further improve the yield of trehalose. The research progress on trehalose biosynthesis pathways and the applic...This paper aims to provide better guidance for construction of trehalose-producing recombinant strains to further improve the yield of trehalose. The research progress on trehalose biosynthesis pathways and the application of molecular biology technique in trehalose study in recent 30 years, especially the last 10 years are reviewed. Results show that there are 5 pathways of trehalose synthesis. Although enzymes and genes of trehalose synthesis have been isolated and genetic engineering strains have increased gradually, the improvement of trehalose yield is still inadequate because most recombinant strains are limited to study the physicochemical properties of single enzyme. With the development of modern biological technology, especially the rapid development of DNA recombinant technology, metagenomics and synthetic biology, high expression of heterologous trehalose in recombinant strains would become a hot research topic in the future.展开更多
The present study is aimed at studying the gene for TIMP-3,a mammalian tissue inhibitor,by constructing a recombinant eukaryotic cell vector for gene therapy in human breast cancer.We obtained the TIMP-3 gene from the...The present study is aimed at studying the gene for TIMP-3,a mammalian tissue inhibitor,by constructing a recombinant eukaryotic cell vector for gene therapy in human breast cancer.We obtained the TIMP-3 gene from the human placent by RT-PCR.TIMP-3 gene was subcloned into pcDNA3.1 vetor from pMD18T vector by means of gene cloning to construct pcDNA3.1 recombinant vector.Human breast cancer cell line MDA-MB-453 was transfected with pcDNA3.1-TIMP3 recombinant vector using lipofectamine reagent.Then the expression of TIMP-3 and the effect on the metastasis of MDA-MB-453 were examined.The correct construction of pcDNA-TIMP3 was identified by means of restriction enzyme analysis,PCR amplication and nucleotide sequencing.Western blotting showed that the transfected cells were able to express TIMP-3, indicating that our construction of the pcDNA-TIMP3 eukaryotic expression vector was constructed successfully.Our experiments further indicated that the potential of metastasis was significantly reduced for the transfected cell line MDA-MB-453.Cellular & Molecular Immunology.2004;1(4):308-310.展开更多
文摘This paper aims to provide better guidance for construction of trehalose-producing recombinant strains to further improve the yield of trehalose. The research progress on trehalose biosynthesis pathways and the application of molecular biology technique in trehalose study in recent 30 years, especially the last 10 years are reviewed. Results show that there are 5 pathways of trehalose synthesis. Although enzymes and genes of trehalose synthesis have been isolated and genetic engineering strains have increased gradually, the improvement of trehalose yield is still inadequate because most recombinant strains are limited to study the physicochemical properties of single enzyme. With the development of modern biological technology, especially the rapid development of DNA recombinant technology, metagenomics and synthetic biology, high expression of heterologous trehalose in recombinant strains would become a hot research topic in the future.
文摘The present study is aimed at studying the gene for TIMP-3,a mammalian tissue inhibitor,by constructing a recombinant eukaryotic cell vector for gene therapy in human breast cancer.We obtained the TIMP-3 gene from the human placent by RT-PCR.TIMP-3 gene was subcloned into pcDNA3.1 vetor from pMD18T vector by means of gene cloning to construct pcDNA3.1 recombinant vector.Human breast cancer cell line MDA-MB-453 was transfected with pcDNA3.1-TIMP3 recombinant vector using lipofectamine reagent.Then the expression of TIMP-3 and the effect on the metastasis of MDA-MB-453 were examined.The correct construction of pcDNA-TIMP3 was identified by means of restriction enzyme analysis,PCR amplication and nucleotide sequencing.Western blotting showed that the transfected cells were able to express TIMP-3, indicating that our construction of the pcDNA-TIMP3 eukaryotic expression vector was constructed successfully.Our experiments further indicated that the potential of metastasis was significantly reduced for the transfected cell line MDA-MB-453.Cellular & Molecular Immunology.2004;1(4):308-310.
