Watanabe T, Kono M, Aida K, Nagasawa H
Laboratory of Molecular Biology of Marine Organisms, University of Tokyo, Japan.
Mol Mar Biol Biotechnol. 1996 Dec;5(4):299-303.
Amino acid sequences of chitinases have been determined in insects, plants, yeast, and bacteria, but not in crustaceans. We searched for chitinase-encoding cDNA in the kuruma prawn Penaeus japonicus by polymerase chain reaction (PCR) amplification of hepatopancreas cDNA using degenerate oligonucleotide primers derived from the two conserved regions of known chitinases. Using a PCR product as a probe, a cDNA clone was isolated. This clone contains an open reading frame for a protein (named Pjchi-1) of 572 amino acids that exhibits sequence similarities to known chitinases, especially to a chitinase from the tobacco hornworm Manduca sexta. Transcription of the mRNA was detected in the hepatopancreas but not in epidermal tissues.
几丁质酶的氨基酸序列已在昆虫、植物、酵母和细菌中得到测定，但尚未在甲壳类动物中测定。我们通过聚合酶链反应（PCR）扩增日本对虾肝胰腺cDNA，使用从已知几丁质酶的两个保守区域衍生而来的简并寡核苷酸引物，在日本对虾（Penaeus japonicus）中寻找编码几丁质酶的cDNA。以PCR产物为探针，分离出一个cDNA克隆。该克隆包含一个572个氨基酸的蛋白质（命名为Pjchi-1）的开放阅读框，该蛋白质与已知的几丁质酶具有序列相似性，尤其是与烟草天蛾（Manduca sexta）的一种几丁质酶相似。在肝胰腺中检测到了mRNA的转录，但在表皮组织中未检测到。
