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Induction of extremely low protein expression level by fusion of C‐terminal region of Nef
Author(s) -
Takamune Nobutoki,
Irisaka Yukari,
Yamamoto Minami,
Harada Keisuke,
Shoji Shozo,
Misumi Shogo
Publication year - 2012
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1021
Subject(s) - fusion protein , mutant , hek 293 cells , biology , protein expression , microbiology and biotechnology , fusion , luciferase , cell fusion , cell culture , chemistry , gene , biochemistry , genetics , recombinant dna , transfection , linguistics , philosophy
Nef is one of the accessory proteins of human immunodeficiency viruses. Here, we noted that the relative expression level of Nef NL4‐3 is much lower than that of Nef JR‐CSF in HEK293 cells. By evaluating the expression level using a Nef mutant, it was indicated that amino acids 129‐206 of Nef NL4‐3 , that is, the C‐terminal region named NLAA129‐206, could contain the region responsible for the induction of the low protein expression level. In addition, the expression levels of the enhanced green fluorescent protein and Renilla luciferase became extremely low with the fusion of NLAA129‐206. Interestingly, the NLAA129‐206‐corresponding sequences of other Nef variants with relatively high expression levels also induced the extremely low protein expression level by fusion. These results suggest that the C‐terminal region of Nef can generally induce an extremely low protein expression level. Here, we propose that the C‐terminal region of Nef could become an excellent tool for the induction of an extremely low expression level of arbitrary proteins by attachment as fusion proteins.