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Permanent amniocyte cell line, the production thereof and its use for producing gene transfer vectors

An amniotic fluid cell, permanent technology, applied in the direction of using vectors to introduce foreign genetic material, vectors, nucleic acid vectors, etc., can solve the problem of not providing information for use

Inactive Publication Date: 2003-07-30
赛维科药物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, transformation of human cells with SV40 is uninformative for transformation with the E1 function of adenovirus, and its use to generate gene transfer vectors

Method used

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  • Permanent amniocyte cell line, the production thereof and its use for producing gene transfer vectors
  • Permanent amniocyte cell line, the production thereof and its use for producing gene transfer vectors
  • Permanent amniocyte cell line, the production thereof and its use for producing gene transfer vectors

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0102] 1. Clone

[0103] Figure 1 depicts a summary of the clones.

[0104] a) Plasmid STK136

[0105] Plasmid STK136, containing the mouse phosphoglycerate kinase promoter (SEQ NO. 1; Adra et al., Gene 60, 65-74, 1987) in [Bluescript KsII (Stratagene), was generated as follows:

[0106] 3.5 micrograms of plasmid PGK-hAAT (Kay et al., Hepatology 21, 815-819, 1995) were digested with EcoRV and fractionated by size on a 1.5% agarose gel. After ethidium bromide staining, look for a 0.5kb band containing the PGK promoter, cut it out, and electroelute the DNA. Simultaneously, pBluescript KSII was digested with EcoRV and HincII to dephosphorylate free DNA ends. Then, after extraction with phenol / chloroform and ethanol precipitation, these DNA fragments were ligated in equimolar amounts and transformed into supercompetent XL-2Blue bacteria (Stratagene). A plasmid clone was characterized by restriction digest and the resulting plasmid was designated STK136 (isolate #6).

[0107] ...

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Abstract

The invention relates to a permanent amniocyte cell line containing at least one nucleic acid, which effects the expression of gene products of the E1A region and E1B region of andenovirus. The invention also relates to the production of a permanent amniocyte cell line and to its use for producing gene transfer vectors. Additional aspects include the use of amniocytes or o f adenoviral gene products of the E1A region and E1B region for producing permanent amniocyte cell lines.

Description

field of invention [0001] The present invention relates to permanent amniotic fluid cell lines containing at least one nucleic acid capable of expressing the gene product of the E1A and E1B regions of an adenovirus. The present invention also relates to the preparation of the permanent amniotic fluid cell line and its use for the preparation of gene transfer vectors and / or adenovirus mutants. A further aspect is the use of amniocytes and adenoviral gene products of the E1A and E1B regions for the production of permanent amniotic cell lines. Background of the invention [0002] Adenovirus [0003] Adenovirus is a relatively uniform virus characterized by an icosahedral capsid mainly composed of virally encoded hexons, pentons and fibrin, and a linear double-stranded DNA genome of about 36 kb in size. The viral genome contains inverted terminal repeats (ITRs) at the ends, which have the viral origin of replication. A packaging signal is also contained at the left end of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N5/10C12N7/00C12N15/00C12N15/861
CPCC12N2810/6018C12N7/00C12N2710/10343C12N2810/405C12N2710/10352C12N2810/60C12N2810/859C12N2810/40C12N2800/30C12N15/86
Inventor G·希德尔史蒂芬·科全耐克
Owner 赛维科药物有限公司