[Study on antifreeze protein in fishes. II. The cloning of antifreeze protein gene cDNA of Pseudopleuronectes yokohamae and its expression in E. coli].
Pseudopleuronectes yokohamae, existing in the Yellow Sea of China, contains antifreeze peptide (AFP). This protein could depress the serum freezing point. On the basis of purification and characterization of this protein, we synthesized a segment of Oligo-nucleotides of antifreeze protein gene as a primer and hybridized with the mRNA of Pseudopleuronectes yokohamae. The cDNA of antifreeze protein gene has specifically been reversely transcribed. This segment has been cloned onto pUC19 with Ecor I linker method. After confirmation of the insert segment of the cDNA of antifreeze protein gene, the zymogram was detected and the sequence of the nucleotides was determined. This recombinated clone was transformed into E. coli JM83 and expressed well.
Jiang
Y Q
YQ
Institute of Genetics, Academia Sinica, Beijing.
Chen
X F
XF
Liu
P T
PT
Zhang
N C
NC
D000595:Amino Acid Sequence; D000818:Animals; D021301:Antifreeze Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004926:Escherichia coli / Q000378:metabolism*; D005399:Fishes / Q000235:genetics*; D005615:Freezing; D006023:Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data
D021301:Antifreeze Proteins; D006023:Glycoproteins; D004247:DNA