甘蓝型油菜β-碳酸酐酶 cDNA 的克隆与表达。

Cloning and expression of Brassica napus beta-carbonic anhydrase cDNA.

机构信息

Institute of Resource Biology & Biotechnology, Department of Biotechnology, College of Life Science & Technology, Huazhong University of Science & Technology, Wuhan 430074, P. R. China.

出版信息

Z Naturforsch C J Biosci. 2009 Nov-Dec;64(11-12):875-81. doi: 10.1515/znc-2009-11-1220.

DOI:10.1515/znc-2009-11-1220

PMID:20158161

Abstract

A new full-length beta-carbonic anhydrase cDNA was obtained from Brassica napus by homologous cloning. The cDNA has an open-reading frame of 996 nucleotides, encoding 331 amino acids with a calculated molecular weight of 35,692 Da and an estimated pI value of 5.459. The deduced amino acid sequence of beta-carbonic anhydrase from Brassica napus shared significant identity with beta-carbonic anhydrases from Brassica carinata, Arabidopsis thaliana, and Thlaspi caerulescens (97.9%, 94%, and 93.5% identity, respectively). This cDNA was expressed in Escherichia coli BL21 (DE3) using the expression vector pET-32a(+). The expression band corresponded to the calculated mass plus the N-terminal fusion protein derived from the vector.

摘要

通过同源克隆，从油菜中获得了一个新的全长β-碳酸酐酶 cDNA。该 cDNA 具有 996 个核苷酸的开放阅读框，编码 331 个氨基酸，计算分子量为 35692Da，估计等电点为 5.459。油菜β-碳酸酐酶的推导氨基酸序列与甘蓝型油菜、拟南芥和蓝蓟的β-碳酸酐酶具有显著的同源性（分别为 97.9%、94%和 93.5%的同一性）。该 cDNA 利用表达载体 pET-32a(+)在大肠杆菌 BL21(DE3)中表达。表达带与计算的质量加上来自载体的 N 端融合蛋白相对应。