cDNA cloning and Escherichia coli expression of UK114 tumor antigen
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- 作者：Colombo ; Irma ; Ceciliani ; Fabrizio ; Ronchi ; Severino ; Bartorelli ; Alberto ; Berra ; Bruno
- 关键词：UK114 ; UK101 ; Tumor antigen ; Recombinant protein ; Nucleotide sequence ; Translational inhibitor
- 刊名：Biochimica et Biophysica Acta (BBA)/Gene Structure and Expression
- 出版年：1998
- 出版时间：October 23, 1998
- 年：1998
- 卷：1442
- 期：1
- 页码：49-59
- 全文大小：450 K
文摘
Experimental evidence indicates that the antineoplastic effects of UK101, a goat liver perchloric acid extract, is likely due to one of its constituent proteins: the 14 kDa protein named UK114. The cDNA encoding UK114, obtained by PCR methodologies, contains an open reading frame coding for a protein of 137 amino acids with a theoretical molecular mass of 14 298 Da. It shows high sequence homology with a 14 kDa protein identified in human, rat and Mus musculus tissues which is likely involved in the inhibition of cell-free protein synthesis. Northern blot analysis indicated that the transcript is present in variable amounts in a wide range of human tissues. Genomic Southern blots revealed that the UK114 mRNA in goat as well as in human is encoded by a single gene, as is the case in rat. The expression system for UK114 was constructed under the control of the PL promoter from bacteriophage λ and the cDNA coding region has been highly expressed in Escherichia coli as a thioredoxin fusion protein. The recombinant UK114, purified to homogeneity, is immunoreactive to rabbit antisera prepared against UK101 or native UK114, as well as to sera of UK101-treated cancer patients. It inhibits cell-free protein synthesis at 8 μM concentration.