CLONING AND EXPRESSION OF A HUMAN INTERFERON a2 GENE IN E. COLI
The plasmid pALCA1SIFN containing cDNA that encodes the human interferon a-2b was obtained from the ATCC(no. 531667). In this system the expression of the gene is under the control of an alcA promoter. alcA p is a specific promoter for expression of different genes in Aspergillusfilamentous. In this...
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| Language: | English |
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University of Tehran
1999-06-01
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| Series: | Journal of Sciences, Islamic Republic of Iran |
| Online Access: | https://jsciences.ut.ac.ir/article_31487_fe2b5abdb5995f963ecedef9efe18921.pdf |
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| description | The plasmid pALCA1SIFN containing cDNA that encodes the human interferon
a-2b was obtained from the ATCC(no. 531667). In this system the expression of the
gene is under the control of an alcA promoter. alcA p is a specific promoter for
expression of different genes in Aspergillusfilamentous. In this plasmid the coding
region of IFN?-2b is preceded by the coding region of a synthetic signal peptide. For
direct expression of the IFNa-2b gene under the control of a T7/lac promotor of
pET24d(+) expression vector, three subcloning steps were carried out which resulted
in the construction of 3 new plamids. pHA1, pHA2 and pHA4 in which the IFNa-2
gene is under the control of lacp, lacuv5p and T7 promoter, respectively. Another
plasrnid, pHA3, was also constiucted and is a modified version of pET24d(+).
Provided there is a synthetic signal peptide preceding the coding region of IFNa2
gene, the protein can be seen in either system, as shown by western blotting, albeit
with a different level of expression. The best product can be seen in the pHA4plasmid
with a T7 pas judged by dot and western blotting |
| format | Article |
| id | doaj-art-7e10b4c0c47d419cbafed11e8fee7e13 |
| institution | OA Journals |
| issn | 1016-1104 2345-6914 |
| language | English |
| publishDate | 1999-06-01 |
| publisher | University of Tehran |
| record_format | Article |
| series | Journal of Sciences, Islamic Republic of Iran |
| spelling | doaj-art-7e10b4c0c47d419cbafed11e8fee7e132025-08-20T01:53:33ZengUniversity of TehranJournal of Sciences, Islamic Republic of Iran1016-11042345-69141999-06-0110231487CLONING AND EXPRESSION OF A HUMAN INTERFERON a2 GENE IN E. COLIThe plasmid pALCA1SIFN containing cDNA that encodes the human interferon a-2b was obtained from the ATCC(no. 531667). In this system the expression of the gene is under the control of an alcA promoter. alcA p is a specific promoter for expression of different genes in Aspergillusfilamentous. In this plasmid the coding region of IFN?-2b is preceded by the coding region of a synthetic signal peptide. For direct expression of the IFNa-2b gene under the control of a T7/lac promotor of pET24d(+) expression vector, three subcloning steps were carried out which resulted in the construction of 3 new plamids. pHA1, pHA2 and pHA4 in which the IFNa-2 gene is under the control of lacp, lacuv5p and T7 promoter, respectively. Another plasrnid, pHA3, was also constiucted and is a modified version of pET24d(+). Provided there is a synthetic signal peptide preceding the coding region of IFNa2 gene, the protein can be seen in either system, as shown by western blotting, albeit with a different level of expression. The best product can be seen in the pHA4plasmid with a T7 pas judged by dot and western blottinghttps://jsciences.ut.ac.ir/article_31487_fe2b5abdb5995f963ecedef9efe18921.pdf |
| spellingShingle | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI Journal of Sciences, Islamic Republic of Iran |
| title | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI |
| title_full | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI |
| title_fullStr | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI |
| title_full_unstemmed | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI |
| title_short | CLONING AND EXPRESSION OF A HUMAN
INTERFERON a2 GENE IN E. COLI |
| title_sort | cloning and expression of a human interferon a2 gene in e coli |
| url | https://jsciences.ut.ac.ir/article_31487_fe2b5abdb5995f963ecedef9efe18921.pdf |