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Expression of human lymphotoxin alpha in Aspergillus niger

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Author: Krasevec, N. · Hondel, C.A.M.J.J. van de · Komel, R.
Type:article
Date:2000
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Pflügers Archiv - European Journal of Physiology, suppl. 5, 440, p. R83-R85
Identifier: 56828
Keywords: Medicine · Geneeskunde · Pathology · Aspergillus niger · Codon usage · Heterologous protein secretion · LTα · TNFα · Complementary DNA · Lymphotoxin · Tumor necrosis factor alpha · Aspergillus niger · Codon usage · Conference paper · Controlled study · DNA sequence · Gene control · Gene fusion · Genetic transcription · Human · Nonhuman · Priority journal · Protein degradation · Protein secretion · Artificial Gene Fusion · Aspergillus niger · Blotting, Northern · Glucan 1,4-alpha-Glucosidase · Humans · Lymphotoxin-alpha · RNA, Messenger

Abstract

A gene-fusion expression strategy was applied for heterologous expression of human lymphotoxin alpha (LTα) in the Aspergillus niger AB1.13 protease-deficient strain. The LTα gene was fused with the A. niger glucoamylase GII-form as a carrier-gene, behind its transcription control and secretion signals. Special attention was paid to the influence of different codon usage on secretion of protein. In the case of human tumor necrosis factor alpha (TNFα) a dramatic change of secretion has been observed when human cDNA sequence was used instead of synthetic E. coli biased codons. In the case of LTα such a change of codon usage brought improvement at the RNA level, however, no increase in the quantity of secreted protein was observed, due to the proteolitic activity of the host organism. The estimated yield of secretion of LTα from A. niger into the soya medium was 50 pg l-1 of culture. Chemicals/CAS: Glucan 1,4-alpha-Glucosidase, EC 3.2.1.3; Lymphotoxin-alpha; RNA, Messenger