Cloning And Bioinformatics Analysis Of Two Genes BnbZIP And BnHSP In Brassica Napus

Stress related genes play a significant role in plant environmental stress.In this study,two stress related genes were cloned from B.napus,which were Leucine zipper protein gene(BnbZIP)and Heat shock protein gene(BnHSP).The sequence characteristics and expression patterns of these two genes were analysed,and their overexpression vectors were constructed and transformed into B.napus,and transformants were obtained.These results laid a foundation for further studies on gene function of BnbZIP and BnHSP in plants.The main results are as follows:1.Leucine zipper protein gene in Brassica napus(BnbZIP)? In this paper,a bZIP gene was cloned in Brassica napus and designated as BnbZIP.The open reading frame(ORF)of BnbZIP is 534 bp in length,and encodes 177 amino acids with a predicted molecular mass of 19875.1 and a pI of 6.91,has a half-life of 30 h,hydrophilic for 0.934.? In total,192 bZIP genes were identified in the genome of Brassica napus.The CDS nucleotide sequences of bZIPs are ranging from 153 to 1908 in length,with an average length of 816 nucleotides.The deduced proteins are ranging from 43 to 601 amino acids in length.? Phylogenetic analysis on bZIP gene families showed a conserved evolutionary pattern,and the deduced BnbZIP protein was closest to bZIP protein in Arabidopsis.? BnbZIP expressed in all tissues analysed,with a highest expression level in 7-day-old hypocotyls,moderately expression in 7-day-old cotyledons,and lowest expression in stems at flowering stage.? We constructed overexpression of BnbZIP plant expression vector,and transformed it into B.napus cultivar "Zhong Shuang 11" by means of Agrobacterium-mediated floral-dip infection,finally 8 positive transformants of T0 plants overexpressing BnbZIP gene were obtained.2.Heat shock protein gene in brassica napus(BnHSP)? In this paper,a HSP gene was cloned in Brassica napus and designated as BnHSP.The open reading frame(ORF)of BnHSP is 2364bp in length,and encodes 787 amino acids with a predicted molecular mass of 89563.9 and a pI of 4.97,has a half-life of 30 h,hydrophilic for 0.593.?In total,87 bZIP genes were identified in the genome of Brassica napus.The CDS nucleotide sequences of HSPs are ranging from 96 to 2925 in length,with an average length of 816 nucleotides.The deduced proteins are ranging from 31 to 973 amino acids in length.? Phylogenetic analysis showed that the deduced BnHSP protein was closest to HSP protein in Brassica rapa.?BnHSP transcript is expressed in all tissues analysed,with highest expression in floral organs,moderately expression in stems and young siliques,and lowest expression in 7-day-old hypocotyls.? We constructed overexpression of BnHSP plant expression vector,and transformed it into B.napus cultivar"Zhong Shuang 11" by means of Agrobacterium-mediated floral-dip infection,finally 10 positive transformants of TO plants overexpressing BnHSP gene were obtained.