CAJ | 학술논문

通过RT-PCR技术从甘薯的总 RNA 中克隆得到了全长的己糖激酶基因,测序结果表明HXK(hexokinase)基因编码区长1491bp,编码496个氨基酸,其翻译的蛋白质分子量为53.4kD,其氨基酸序列与其它已知高等植物HXK基因具有很高的同源性.构建了原核表达载体pET-HXK,经IPTG诱导后可表达获得相对分子量约为71 kD的外源融合蛋白,与预测结果一致.并对HXK基因在不同组织中的转录水平进行了数字表达谱和荧光定量分析.
통과RT-PCR기술종감서적총 RNA 중극륭득도료전장적기당격매기인,측서결과표명HXK(hexokinase)기인편마구장1491bp,편마496개안기산,기번역적단백질분자량위53.4kD,기안기산서렬여기타이지고등식물HXK기인구유흔고적동원성.구건료원핵표체재체pET-HXK,경IPTG유도후가표체획득상대분자량약위71 kD적외원융합단백,여예측결과일치.병대HXK기인재불동조직중적전록수평진행료수자표체보화형광정량분석.
The HXK gene in sweet potato contains an open reading frame (ORF)of 1,491 bp encoding a peptide of 496 amino acids with a molecular mass of 53.4 kD.The amino acid sequence of IbHXK has high homology with HXK genes from other plants.The IbHXK gene was also inserted into the pET32a (+)vector,and the recombinant plasmid pET-HXK was then transformated into E.coli BL2 1 (DE3 ) for prokaryotic expression induced by different concentrations of IPTG.The transcript levels of the HXK gene in different tissues were invistigated by the digital gene expression profiling (DGE)and quantitative RT-PCR analysis.