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BANCR (BRAF-activated non-protein coding RNA) overexpressed human skin melanoma stable cell strain and preparation method and application thereof

A melanoma cell and melanoma technology, applied in the field of BANCR overexpression human skin melanoma stable cell line and its preparation, can solve the problem of no melanoma exploration and research

Inactive Publication Date: 2017-07-21
KUNMING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, in the current domestic and foreign reports on the correlation between BANCR and melanoma, there is no BANCR overexpressed human A375 stable transfected cell line as an experimental system for melanoma research.

Method used

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  • BANCR (BRAF-activated non-protein coding RNA) overexpressed human skin melanoma stable cell strain and preparation method and application thereof
  • BANCR (BRAF-activated non-protein coding RNA) overexpressed human skin melanoma stable cell strain and preparation method and application thereof
  • BANCR (BRAF-activated non-protein coding RNA) overexpressed human skin melanoma stable cell strain and preparation method and application thereof

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Embodiment 1

[0046] Example 1 Construction method of BANCR overexpression type human skin melanoma stable cell line

[0047] 1. Synthesis of the full sequence of the human LncRNA BANCR gene

[0048] Select the physical map of LV5 (LV5 plasmid as figure 1 Shown) as an overexpression vector, design the PCR primers required for the synthesis of the human LncRNABANCR gene sequence (SEQ ID NO: 7), design by oligo online software (http: / / www.oligo.net / ), and add upstream and downstream primers respectively Homologous sequences on both sides of Not I and Nsi I on the LV5 vector are used for subcloning of the vector (primers are synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd.), and the designed primers are as follows:

[0049] BANCR-F:

[0050] 5'-AGGGTTCCAAGCTTAAGCGGCCGCATTCCCTTACTTTCTTAATAAAC-3' (SEQ ID NO: 3)

[0051] BANCR-R:

[0052] 5'-GATCCATCCCTAGGTAGATGCATTTTTTTTTTTTTTAGGATTTTTTA-3' (SEQ ID NO: 4)

[0053] After diluting the synthesized primers to 50 μM, carry out P...

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Abstract

The invention provides a BANCR (BRAF-activated non-protein coding RNA) overexpressed human skin melanoma stable cell strain and a preparation method and application thereof and relates to the technical field of molecular biology. BANCR gene is inserted into an expression vector to obtain a recombinant lentiviral vector, packaging cells are co-transfected by the recombinant lentiviral vector and packaging plasmid to obtain recombinant lentivirus, human skin melanoma cells are infected with the recombinant lentivirus to obtain the BANCR overexpressed human skin melanoma stable cell strain. The construction method provided herein integrates a target gene to a host cell through a lentivirus system, and finally acquires the efficient stable BANCR overexpressed human skin melanoma stable cell strain by screening and identifying; the cell strain is used for studying BANCR and tumor relevancy and its mechanism, and reliable study basis is provided for the deep study on possible occurrence and developing mechanisms of BANCR and human melanoma.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a BANCR overexpression type human skin melanoma stable cell line and a preparation method and application thereof. Background technique [0002] The "central dogma" of genetics describes the information flow pathway of genetic information from DNA through RNA to corresponding proteins. For decades, the conventional wisdom has been that DNA is the storage carrier of genetic material, and that genes function through the corresponding proteins they express. With the successful completion of the Human Genome Project, it was discovered that only a small portion of genes are protein-coding genes, while more than 90% of human genes have been transcribed, which indicates that a large number of transcripts do not code for proteins. This is called "non-coding RNA" (non-coding RNA, ncRNA). At first, ncRNA was considered as transcriptional "noise". However, in recent years, more ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867C12Q1/02A01K67/027
CPCA01K67/0278A01K2267/0331C12N5/0693C12N15/1135C12N15/86C12N2740/15043G01N33/5011G01N2500/10
Inventor 朱月春杨丽娟杨雨叶杨慧鑫蒋露白宏刚张巧易子寒
Owner KUNMING MEDICAL UNIVERSITY
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