Biomaterial Database

[Gene cloning and expression of TNF in Streptomyces lividans TK54]. 1993

J Liu, and Y Li, and B Liu

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.

With the plasmid pIJ486 as a vector, TNF cDNA originated from plasmid pHT1 has been cloned into Streptomyces lividans TK54. After transformation, hundreds of transformants, resistant to neomycin (30 micrograms/ml) were selected. Result showed that TNF cDNA was inserted into the plasmid pIJT7 harboured in No. 7 transformant, namely Streptomyces lividans TK54-HT. Cytotoxic activity experiment of L929 cell demonstrated that the maximal TNF activity in Streptomyces lividans TK54-HT is more than 10(8) unit/L culture. The neutralization experiment showed that the product producing by Streptomyces lividans TK54-HT is human TNF-alpha. SDS-PAGE confirmed that there is a 17,000 dolton protein band which has the same molecular weight as that of TNF in the clear lysate supernatant of Streptomyces lividans TK54-HT cells. All of these results mentioned above showed that the TNF gene has been successfully cloned and expressed in Streptomyces lividans TK54.

UI	MeSH Term	Description	Entries
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D013302	Streptomyces	A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.
D014409	Tumor Necrosis Factor-alpha	Serum glycoprotein produced by activated MACROPHAGES and other mammalian MONONUCLEAR LEUKOCYTES. It has necrotizing activity against tumor cell lines and increases ability to reject tumor transplants. Also known as TNF-alpha, it is only 30% homologous to TNF-beta (LYMPHOTOXIN), but they share TNF RECEPTORS.	Cachectin,TNF-alpha,Tumor Necrosis Factor Ligand Superfamily Member 2,Cachectin-Tumor Necrosis Factor,TNF Superfamily, Member 2,TNFalpha,Tumor Necrosis Factor,Cachectin Tumor Necrosis Factor,Tumor Necrosis Factor alpha
D015870	Gene Expression	The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.	Expression, Gene,Expressions, Gene,Gene Expressions