Cloning and expression of Taxus acyltransferase cDNA.
A new full-length acyltransferase cDNA was obtained from Taxus chinensis by homology-based cloning strategy. The cDNA has an open-reading frame of 1,275 nucleotides, which encodes 425 amino acids with a calculated molecular weight of 47,241 Da and an estimated pI value of 5.93. The deduced amino acid sequence resembles the sequences of other cloned acyltransferases (56-61% identity; 71-75% similarity) involved directly in taxol biosynthetic pathways. This cDNA was expressed in Escherichia coli using the expression vector pET32a(+). The expression band corresponds to the calculated mass plus the N-terminal fusion protein derived from the vector.
Tu
Jun
J
Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, P. R. China.
Zhu
Ping
P
Cheng
Ke-Di
KD
Liu
Jiangguo
J
D000217:Acyltransferases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D020946:Taxus / Q000201:enzymology*
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D011994:Recombinant Proteins; D000217:Acyltransferases