摘要
为进一步研究甘蓝型油菜BnaLPAT2基因,通过TAIR网站查找拟南芥AtLPAT2 (AT3G57650),在Brassica Database数据库检索到4条甘蓝型油菜LPAT2序列,据此设计引物,通过RT-PCR合成c DNA,得到4条CDS序列,其大小依次为1 173 bp、1 167 bp、1 173 bp、1 155 bp,分别命名为BnaLPAT2-1~BnaLPAT2-4。4个BnaLPAT2蛋白通过生物信息学预测分析显示:BnaLPAT2编码的氨基酸在384~390之间,理论分子量介于43 226.59~43 730.17 Da之间。甘蓝型油菜LPAT2蛋白的二级结构和跨膜区大致相同,均由LPLAT superfamily、PlsC superfamily、Acyltransf_C superfamily结构域组成,且具有较近的亲缘关系。从基因结构发现:4个BnaLPAT2均含有相同的外显子和内含子数,分别位于C08、A09、A07和A04染色体上。分析表明,克隆获得的4个甘蓝型油菜BnaLPAT2为该基因家族成员。为进一步研究甘蓝型油菜LPAT2基因的功能,我们通过双酶切技术,构建pBI121-LPAT2-napin载体,并转化根癌农杆菌GV3101。
In order to further study BnaLPAT2 in Brassica napus, the Arabidopsis thaliana AtLPAT2 gene(AT3 G57650) was searched on TAIR website and the LPAT2 sequence of Brassica napus was retrieved from the gene of AtLPAT2 and downloaded from GenBank. According to these sequences information, four pairs of primers were designed to amplify four CDS sequences. The length were 1 173 bp, 1 167 bp, 1 173 bp and 1 155 bp, they were named BnaLPAT2-1~BnaLPAT2-4. The protein encoded by BnaLPAT2 in Brassica napus was predicted by bioinformatics software. The results showed that the amino acids encoded by BnaLPAT2 was between 384 and390, and the theoretical molecular weight was between 43 226.59 and 43 730.17 Da. The secondary structure and transmembrane region of BnaL PAT2 protein in Brassica napus L. are roughly the same, containing LPLAT superfamily, PlsC superfamily and Acyltransf_C superfamily domains, and have close genetic relationship. The gene structure map shows that BnaLPAT2 contains the same exon and intron number, and spreads across four different chromosomes. These results showed that BnaLPAT2-1~4 are the four members of the BnaLPAT2 family.In order to further study the function of LPAT2 gene in Brassica napus, in addition, the BnaLPAT2 gene was successfully ligated into the plant binary expression vector p BI121 by double digestion, and transformed into GV3101.
引文
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