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- Title
Changes of primary sequence and secondary structure proximal to the 5' end of the stop codon substantially increases the expression of the variable region of an antibody inE. coli.
- Authors
Weisong Qin; Jiannan Feng; Yan Li; Zhou Lin; Beifen Shen
- Abstract
The sequence context at the 5' end of the stop codon may influence the efficiency of termination and translation. To increase the expression of a designed variable region of an antibody (named as VH5) against tumor necrosis factora(TNFa), two nucleotides (TC) at 25 and 26 nucleotides (nt) upstream of termination codon were substituted with AG, respectively. The free energy of 70 nt (arbitrarily defined from the 32 nt upstream of termination codon to 38 nt downstream) was changed from -13.5 kcal mol-1 to -17.3 kcal mol-1. The expression level was increased from 1 ± 0.3% to 10 ± 1.2% of total cellular protein. Although the precise mechanism of this phenomenon remains to be elucidated, this report provides an alternative means to increase the expression of a foreign gene inE. coli.
- Subjects
ESCHERICHIA coli; TUMOR necrosis factors; IMMUNOGLOBULINS; NUCLEOTIDES; ESCHERICHIA; PROTEOMICS; GLYCOPROTEINS; BIOTECHNOLOGY
- Publication
Biotechnology Letters, 2005, Vol 27, Issue 2, p131
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-004-7660-4