CAJ | 학술논문

CRABS CLAW( CRC)转录因子是YABBY基因家族成员,在植物花器官发育过程中起重要作用。为进一步研究CRC转录因子在调控油菜花器官发育过程中的作用,本研究以甘蓝型油菜品种宁油10号花蕾RNA为材料,通过反转录PCR克隆到1个580 bp长度的CRC基因,并利用pHurricane中间载体构建CRC基因的反向重复序列表达框,将CRC基因片段以正向的方式连接在1个可剪切的内含子的5忆末端,以反向的方式连接到该内含子的3忆末端；然后将CaMV35S启动子序列和CRC基因的反向重复表达框再克隆到植物双元载体 pCAMBIAI1390的pUC18多克隆位点,构建了干扰表达载体pA6-CRCi,经过酶切鉴定和测序分析,所构建的载体正确。
CRABS CLAW( CRC)전록인자시YABBY기인가족성원,재식물화기관발육과정중기중요작용。위진일보연구CRC전록인자재조공유채화기관발육과정중적작용,본연구이감람형유채품충저유10호화뢰RNA위재료,통과반전록PCR극륭도1개580 bp장도적CRC기인,병이용pHurricane중간재체구건CRC기인적반향중복서렬표체광,장CRC기인편단이정향적방식련접재1개가전절적내함자적5억말단,이반향적방식련접도해내함자적3억말단；연후장CaMV35S계동자서렬화CRC기인적반향중복표체광재극륭도식물쌍원재체 pCAMBIAI1390적pUC18다극륭위점,구건료간우표체재체pA6-CRCi,경과매절감정화측서분석,소구건적재체정학。
CRABS CLAW ( CRC) is a member of the YABBYA transcription factor gene family and plays an impor-tant role in the development of plant flower organ. In order to study the CRC transcription factor’ s function in regulating the rape flower organ development, a 580-bp segment was cloned by RT-PCR from the total RNA of rapeseed flower organ. An inverted repeated expression cassette was firstly constructed by cloning the 580-bp segment of CRC gene into an intermediate pHurrican vector. The fragment was then ligated to a spliceable fad2 intron sequence. Finally, both the inverted repeat cas-sette and CaMV35S promoter were inserted into the multiple clone site of binary vector pCAMBIA1390. The ihpRNA ex-pression vector was named pA6-CRCi, and the construction of which was further confirmed by the digestion of restriction enzymes and sequencing.