| 产品名称： | pCD2 |
|---|---|
| 商品货号： | TS144732 |
| Designations： | pCD2 |
| GenBank Number： | X63656 |
| Species： | Escherichia coli (Migula) Castellani and Chalmers |
| Depositors： | The United States of America, RM Blaese, CA Mullen, The United States of America |
| Applications： | in suitable host, produces protein cytosine deaminase |
| Vector： | Construct size (kb): 7.400000095367432 |
| Insert： | DNA: genomic Insert lengths(kb): 1.700000047683716 Gene product: cytosine deaminase codA Target Gene: cytosine deaminase |
| Insert Size (kb)： | 1.700 |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--7.4; BamHI--7.4; EcoRI/BamHI--5.7, 1.7. The insert contains the following restriction sites (approximate kb from the 5 end): PstI--0.09; ClaI--0.67, 1.28; AccI--0.89. Murine fibroblast cell lines such as NIH 3T3 and PA317, transformed with pCD2, express cytosine deaminase. If transformed cells are grown in the presence of non-toxic 5-flurocytosine (5FC), the cytosine deaminase will convert the 5FC into the lethal metabolite 5-fluoruracil which inhibits DNA and RNA synthesis. This cytosine deaminase negative selection system (CDNSS) could prove useful in selectively controlling gene therapy experiments. PA317/pCD2 transformed cell lines expressed 16.3U of cytosine deaminase while NIH 3T3/pCD2 transformed cell lines expressed 3.6U (1U = 1pmol of uracil converted from cytosine/10+E08 cells/minute). Recombinant cytosine deaminase is expressed poorly in eukaryotic cells if 88 bp 5 to naturally occurring GTG start site is present. pCD2 was engineered to remove these 88 bp and convert the GTG start to ATG. |
| Classification： | Enterobacteriaceae, Escherichia |
| References： | Mullen CA, Blaese RM. Cytosine deaminase negative selection system for gene transfer techniques and therapies. US Patent 5,358,866 dated Oct 25 1994 Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796 |
| Disclosure： | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
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| 产品名称： | pCD2 |
|---|---|
| 商品货号： | TS144732 |
| Designations： | pCD2 |
| GenBank Number： | X63656 |
| Species： | Escherichia coli (Migula) Castellani and Chalmers |
| Depositors： | The United States of America, RM Blaese, CA Mullen, The United States of America |
| Applications： | in suitable host, produces protein cytosine deaminase |
| Vector： | Construct size (kb): 7.400000095367432 |
| Insert： | DNA: genomic Insert lengths(kb): 1.700000047683716 Gene product: cytosine deaminase codA Target Gene: cytosine deaminase |
| Insert Size (kb)： | 1.700 |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--7.4; BamHI--7.4; EcoRI/BamHI--5.7, 1.7. The insert contains the following restriction sites (approximate kb from the 5 end): PstI--0.09; ClaI--0.67, 1.28; AccI--0.89. Murine fibroblast cell lines such as NIH 3T3 and PA317, transformed with pCD2, express cytosine deaminase. If transformed cells are grown in the presence of non-toxic 5-flurocytosine (5FC), the cytosine deaminase will convert the 5FC into the lethal metabolite 5-fluoruracil which inhibits DNA and RNA synthesis. This cytosine deaminase negative selection system (CDNSS) could prove useful in selectively controlling gene therapy experiments. PA317/pCD2 transformed cell lines expressed 16.3U of cytosine deaminase while NIH 3T3/pCD2 transformed cell lines expressed 3.6U (1U = 1pmol of uracil converted from cytosine/10+E08 cells/minute). Recombinant cytosine deaminase is expressed poorly in eukaryotic cells if 88 bp 5 to naturally occurring GTG start site is present. pCD2 was engineered to remove these 88 bp and convert the GTG start to ATG. |
| Classification： | Enterobacteriaceae, Escherichia |
| References： | Mullen CA, Blaese RM. Cytosine deaminase negative selection system for gene transfer techniques and therapies. US Patent 5,358,866 dated Oct 25 1994 Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796 |
| Disclosure： | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
