Construction of 18-trisomy syndrome cell model and cell bank of 18-trisomy syndrome cell by SV40LT gene transfection
The invention relates to construction of an 18-trisomy syndrome cell model and a cell bank of 18-trisomy syndrome cell by SV40LT gene transfection for the field of medical research. The construction is mainly characterized in that an SV40LTag-pcDNA3.1(-) recombinant is constructed through a T4DNA li...
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creator | LIU BEI CHEN YAN WENG BINGHUAN HUANG HEFENG CHEN SONGCHANG LI XIAO |
description | The invention relates to construction of an 18-trisomy syndrome cell model and a cell bank of 18-trisomy syndrome cell by SV40LT gene transfection for the field of medical research. The construction is mainly characterized in that an SV40LTag-pcDNA3.1(-) recombinant is constructed through a T4DNA ligase, BamHI, pcDNA3.1(-) DNA and SV40LTag DNA as a matter of routine, the recombinant is purified through competent escherichia coli, the recombinant is introduced into an in-vitro passage cell digested through collagenase II or an 18-trisomy syndrome tissue cell which is logarithmically grown through a lipoid transfection method to be integrated with cell DNA, and a cloned cell which is screened through G418 and contains a positive recombinant is subjected to amplification culture; and a cell which accords with the characteristics of an immortalized cell and is the same as or similar to a primary cell on a cell morphology, a growth curve, a chromosome karyotype, soft agar colony growth, a nude mouse tumorigenesis test, SV40 large T gene detection of transfection cell DNA, an mRNA expression product measurement result and a DNA sequence measurement result is screened as the 18-trisomy syndrome in-vitro research cell model and cryopreserved in liquid nitrogen, so that the foundation is laid for researching the pathogenesis of 18-trisomy syndrome for a long time from cell level in vitro. |
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subjects | BEER BIOCHEMISTRY CHEMISTRY COMBINATORIAL CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE |
title | Construction of 18-trisomy syndrome cell model and cell bank of 18-trisomy syndrome cell by SV40LT gene transfection |
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