Molecular Cloning and Characterization of a Putative BnHEC3 Gene in Oilseed Rape (Brassica Napus)

Xiaoli Tan, Lili Zhang, Zongwei Xia


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.

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