CAJ | 학술논문

根据已知的拟南芥甜菜碱醛脱氢酶基因(BADH1)序列设计特异性引物,利用RT-PCR技术从甘蓝型油菜中克隆出油菜甜菜碱醛脱氢酶基因(BnBADH-1)全长.测序和分析结果表明,BnBADH-1的cDNA全长1506 bp,编码一个包含501个氨基酸残基、分子量为55 kD、等电点(pI)为5.16的假定蛋白质;核酸序列和氨基酸序列与拟南芥的同源性分别为90.24%和93.41%.利用片段两端的粘性末端酶切位点BamHⅠ和SacⅠ将cDNA片段连接在真核表达载体pBI121上;PCR鉴定表明,过量表达载体pBI121-BnBADH-1已成功构建,为下一步的基因功能分析做好了准备.
근거이지적의남개첨채감철탈경매기인(BADH1)서렬설계특이성인물,이용RT-PCR기술종감람형유채중극륭출유채첨채감철탈경매기인(BnBADH-1)전장.측서화분석결과표명,BnBADH-1적cDNA전장1506 bp,편마일개포함501개안기산잔기、분자량위55 kD、등전점(pI)위5.16적가정단백질;핵산서렬화안기산서렬여의남개적동원성분별위90.24%화93.41%.이용편단량단적점성말단매절위점BamHⅠ화SacⅠ장cDNA편단련접재진핵표체재체pBI121상;PCR감정표명,과량표체재체pBI121-BnBADH-1이성공구건,위하일보적기인공능분석주호료준비.
The full length of cDNA of betaine aldehyde dehydrogenase gene (BnBADH-1) was cloned from Brassica napus by using the specific primers designed according to the sequence of BADH1 from Arabidopsis thaliana. The sequencing and analysis showed that the full length cDNA contained 1 506 bp in length and the deduced amino acid sequence contained 501 amino acid residues, which shared 90.24%and 93.41%identity with the nucleotide sequence and amino acid sequence of betaine aldehyde dehydrogenase gene from Arabidopsis thaliana, respectively;the molecular weight of the protein was 55 kD, and its isoelectric point (pI) was 5.16. The eukaryotic expression vector was constructed using the BamHⅠand SacⅠsites, which would be used in functional analysis in the future research.