Construction of chromosome deletion cat cry syndrome cell model by virtue of SV40LT recombination and cell bank of chromosome deletion cat cry syndrome cell model

Abstract

The invention relates to construction of an SV40LT antigen gene-mediated chromosome deletion cat cry syndrome cell model and a cell bank of the SV40LT antigen gene-mediated chromosome deletion cat cry syndrome cell model in the medical field. The construction is mainly characterized by constructing an SV40LTag-pcDNA3.1 (-) recon by T4DNA ligase, BamHI, pcDNA3.1 (-) DNA and SV40LTag DNA according to a conventional way, purifying the recon by competent escherichia coli, and introducing the purifed recon into chromosome deletion cat cry syndrome cells which are digested by collagenase II and are in in-vitro passage or logarithmic growth by a liposome transfection method, thereby integrating the recon with DNA of the cells, performing amplification culture on cell clones which contain the positive recon and are screened by G418, screening in the aspects of cell morphology, growth curve, karyotype and soft-agar colony growth, performing carcinogenicity testing on nude mice, detecting SV40 large T genes in the DNA of the transfected cells, determining an mRNA expression product and determining a DNA sequence, wherein the cells with the results which are in line with the properties of immortalized cells and are the same with or similar to primary culture cells are taken as the cell model for in-vitro research of SV40LT-mediated chromosome deletion cat cry syndrome to be cryopreserved in liquid nitrogen, and a foundation is further laid for long-term in-vitro research of pathogenesis on cell level.

Description

technical field [0001] The invention relates to SV40LT (simian kidney virus 40 large T antigen gene or SV40LT antigen gene) recombination to construct a chromosomal deletion meowing syndrome cell model and its cell bank, which is mainly used in the field of medical cytogenetics research, and is a chromosomal deletion meowing syndrome The research provides cell models and preserves its scientific research resources. Background technique [0002] Chromosomal deletion cat call syndrome, also known as cri-du-chat syndrome, is a special disease first reported by French scientists in 1963. Cat-calling syndrome in children is caused by the deletion of the short arm of chromosome 5, and most of the deletions are the result of two breaks. Theoretically, there are at least four reasons for partial deletion of chromosomes: terminal deletion, middle deletion, translocation and short arm It was found that the missing part has 5p14, so 5p14 is considered to be the characteristic area of ...

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Problems solved by technology

[0009] However, there has been no literature report on the construction of an immortal cell model and its cell bank that can be used to study the pathogenesis of chromosomal deletion meowing syndrome from the cellular level in vitro, and it is impossible to carry out related research projects

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