CAJ | 학술논문

以缩减杂交的原理和SMART cDNA合成方法为基础,开发了一种全长cDNA缩减杂交方法.该方法从微量的2个RNA样品合成全长cDNA,经PCR扩增和在两端连接不同的接头,通过2次缩减杂交获得含有高度浓缩的差异表达的全长或较大的基因片段,克隆后筛选出差异表达的基因.以噻菌灵(Probenazole,PBZ)为诱导剂处理水稻,获得了2个上调表达和1个下调表达基因的较长的cDNA片段.
이축감잡교적원리화SMART cDNA합성방법위기출,개발료일충전장cDNA축감잡교방법.해방법종미량적2개RNA양품합성전장cDNA,경PCR확증화재량단련접불동적접두,통과2차축감잡교획득함유고도농축적차이표체적전장혹교대적기인편단,극륭후사선출차이표체적기인.이새균령(Probenazole,PBZ)위유도제처리수도,획득료2개상조표체화1개하조표체기인적교장적cDNA편단.
cDNA suppression subtractive hybridization (SSH) has been used to analyze differential gene expression widely, but there are some shortcomings in it such as difficult mRNA preparation, difficult mRNA handling and short cDNA fragments to be obtained.A Full-length cDNA SSH (FLSSH) method based on SSH and SMART cDNA synthesis method was used to obtain differentially-expressed genes of rice treated by probenazole(PBZ).By northern blot analysis, two up-regulated and one down-regulated candidate cDNA fragments by PBZ were identified.