Molecular cloning of a cDNA clone for tobacco lipid transfer protein and expression of the functional protein in Escherichia coli

FEBS Letters

Furuno, Masahiro²  Tanaka, Hideo²  Koiwai, Akira²  Masuta, Chikara²  Yamada, Mitsuhiro¹ 

[1] Department of Bioscience and Technology, School of Engineering, Hokkaido Tokai University, Minamisawa 5-1, Minami-ku, Sapporo 005, Japan;Life Science Research Laboratory, Japan Tobacco Inc., Midori-ku, Yokohama 227, Japan

关键词: Lipid transfer protein;    cDNA;    DNA expression;    Escherichia coli;    Tobacco;   

DOI  :  10.1016/0014-5793(92)81381-U

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A cDNA clone encoding a lipid transfer protein (LTP) was isolated from tobacco by screening a library with a PCR-amplified spinach LTP gene, DNA sequence analysis showed a large open reading frame (344 bp) encoding a polypeptide of 114 amino acids. The first 23 amino acids of the deduced protein have the characteristics of a signal peptide for protein secretion or targeting into dense microbody-like vesicles. The cDNA clone was then inserted into an expression vector, pMAL, and expressed in E. coli as a fusion with the maltose binding protein (MBP). The MBP-LTP fusion protein was purified to homogeneity and subjected to factor Xa cleavage to yield the LTP domain. A lipid transfer assay demonstrated that the resulting LTP was functional. The availability of the expression system in E. coli will facilitate the elucidation of in vivo function(s) of plant LTPs.

【 授权许可】

Unknown