Cloning of trichosanthin cDNA and its expression in Escherichia coli.
Several cDNA clones coding for trichosanthin (TCS) have been isolated from a cDNA library prepared from the poly(A)+RNA of the root tuber of Trichosanthes kirilowii Maximowicz. The nucleotide sequence codes for a protein of 289 amino acids (aa) including a putative signal peptide of 23 aa and an extra 19 aa at the C terminus; the latter two have not been found in TCS obtained from the natural product [Collins et al., J. Biol. Chem. 265 (1990) 8665-8669]. Recombinant TCS (reTCS) was synthesized in Escherichia coli, in which the cDNA without the signal sequence was expressed under the control of the trc promoter; reTCS was detected by a rabbit anti-TCS antiserum.
Shaw
P C
PC
Department of Biochemistry, Chinese University of Hong Kong, Shatin, N.T.
Yung
M H
MH
Zhu
R H
RH
Ho
W K
WK
Ng
T B
TB
Yeung
H W
HW
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D015978:Trichosanthin / Q000096:biosynthesis / Q000235:genetics*
D011994:Recombinant Proteins; D015978:Trichosanthin; D004247:DNA