Cloning and expression of Brassica napus beta-carbonic anhydrase cDNA.
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.
Deng
Qiu-Hong
QH
Institute of Resource Biology & Biotechnology, Department of Biotechnology, College of Life Science & Technology, Huazhong University of Science & Technology, Wuhan 430074, P. R. China.
Li
Mao-Teng
MT
Yu
Long-Jiang
LJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D029688:Brassica napus / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D002256:Carbonic Anhydrases / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D010802:Phylogeny; D010944:Plants / Q000145:classification / Q000201:enzymology / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D002256:Carbonic Anhydrases