苏云金芽孢杆菌达科他亚种中两个新型晶体蛋白基因的克隆与特性分析

Cloning and characterization of two novel crystal protein genes from a Bacillus thuringiensis serovar dakota strain.

作者信息

Kim Ho San, Saitoh Hiroyuki, Yamashita Satoko, Akao Tetsuyuki, Park Yu Shin, Maeda Minoru, Tanaka Rie, Mizuki Eiichi, Ohba Michio

机构信息

Biotechnology & Food Research Institute, Fukuoka Industrial Technology Center, Aikawa-machi 1465-5 Kurume, Fukuoka 839-0861, Japan.

出版信息

Curr Microbiol. 2003 Jan;46(1):33-8. doi: 10.1007/s00284-002-3801-y.

DOI:10.1007/s00284-002-3801-y

PMID:12432461

Abstract

Two genes encoding the 32- and 40-kDa polypeptides of Bacillus thuringiensis strain 90-F-45-14 crystals were cloned, expressed in an acrystalliferous B. thuringiensis strain, and sequenced. The polypeptides had deduced molecular weights of 30,319 and 33,885, respectively. The amino acid sequence of the 32-kDa protein was 37.7% identical to the known sequence of a non-insecticidal parasporal protein in B. thuringiensis serovar thompsoni crystals. The sequence of the cloned 40-kDa protein was 37.0% and 22.3% identical to that of the existing Cry protein classes, Cry15Aa1 and Cry23Aa1, respectively. Thus, this protein constitutes a novel protein class, Cry33Aa1. The open reading frames of the two genes were located on the predominant plasmid of 17,629 bp (=11,752 MDa) in the same orientation, and they were separated by the sequence of 32 nucleotides. The two proteins are likely produced simultaneously from a single transcript to form spherical crystals.

摘要

克隆了编码苏云金芽孢杆菌90-F-45-14菌株晶体中32 kDa和40 kDa多肽的两个基因，在无晶体的苏云金芽孢杆菌菌株中表达并进行了测序。这两种多肽的推导分子量分别为30,319和33,885。32 kDa蛋白的氨基酸序列与苏云金芽孢杆菌汤普森亚种晶体中一种非杀虫伴孢晶体蛋白的已知序列有37.7%的同一性。克隆的40 kDa蛋白的序列与现有Cry蛋白类Cry15Aa1和Cry23Aa1的序列分别有37.0%和22.3%的同一性。因此，这种蛋白构成了一个新的蛋白类，即Cry33Aa1。这两个基因的开放阅读框位于17,629 bp（=11,752 MDa）的主要质粒上，方向相同，它们被32个核苷酸的序列隔开。这两种蛋白可能从单个转录本同时产生，形成球形晶体。