Open AccessMolecular Cloning and Characterization of a Putative BnHEC3 Gene in Oilseed Rape (Brassica Napus)
Author(s) -
Xiaoxiao Tan,
Lili Zhang,
Zongwei Xia
Publication year - 2009
Publication title -
international journal of biology/internationa journal of biology
Language(s) - English
Resource type - Journals
eISSN - 1916-968X
pISSN - 1916-9671
DOI - 10.5539/ijb.v1n2p71
Subject(s) - brassica , open reading frame , complementary dna , biology , gene , arabidopsis , arabidopsis thaliana , genetics , cloning (programming) , in silico , rapid amplification of cdna ends , coding region , molecular cloning , genome , homologous chromosome , microbiology and biotechnology , peptide sequence , botany , mutant , computer science , programming language
HECATE3 (HEC3) is an important transcription factor involved in regulating the carpel and transmitting tract development in Arabidopsis thaliana. The homologous gene of HEC3 in Brassica napus (designed as BnHEC3) was obtained by in silico cloning and rapid amplification of cDNA end (RACE) method. BnHEC3 obtained from cDNA is of 748 bp, containing an open reading frame (ORF) of 546bp and coding a peptide of 181 amino acid residues. Sequence alignment revealed that BnHEC3 is highly homologous to HEC3 in Arabidopsis thaliana, especially in bHLH region, which is sharing nearly 100% identity, and the polygenetic analysis indicated that BnHEC3 belongs to atypical bHLH protein family. RT-PCR results showed that BnHEC3 is mainly expressed in young leave. Southern blotting analysis indicated that BnHEC3 has at least three copies in Brassica napus genome. The transient expression of BnHEC3-sGFP in onion epidermic cells showed that BnHEC3 localized in nuclei. These features implied that BnHEC3 functions as HEC3, and may be involved in the development of female reproductive tissues in Brassica napus