CAJ | 학술논문

　　隐花色素CRY1在拟南芥光形态建成及光信号转导中起重要作用，在津田芜菁(Brassica rapa L. subsp. rapa‘Tsuda’)中是否起相同作用尚未报道。本研究利用实验室已克隆得到的津田芜菁CRY1基因的cDNA全长序列，在NCBI进行序列比对，选取特异的片段。同时根据Gateway克隆技术特点，设计含有attB接头的引物。利用高保真的LA Taq DNA聚合酶，通过PCR方法在目的片段的两端加上attB序列。通过BP反应和LR反应将CRY1基因片段克隆到pH7GWIWG2(I)双元干涉载体。对重组载体pH7GWIWG2(I)-CRY1的鉴定结果表明成功构建了CRY1基因的干涉载体。利用Gateway克隆技术构建植物干涉表达载体简便易行，该结果为遗传转化研究其功能奠定了基础。
　　은화색소CRY1재의남개광형태건성급광신호전도중기중요작용，재진전무정(Brassica rapa L. subsp. rapa‘Tsuda’)중시부기상동작용상미보도。본연구이용실험실이극륭득도적진전무정CRY1기인적cDNA전장서렬，재NCBI진행서렬비대，선취특이적편단。동시근거Gateway극륭기술특점，설계함유attB접두적인물。이용고보진적LA Taq DNA취합매，통과PCR방법재목적편단적량단가상attB서렬。통과BP반응화LR반응장CRY1기인편단극륭도pH7GWIWG2(I)쌍원간섭재체。대중조재체pH7GWIWG2(I)-CRY1적감정결과표명성공구건료CRY1기인적간섭재체。이용Gateway극륭기술구건식물간섭표체재체간편역행，해결과위유전전화연구기공능전정료기출。
The cryptochrome CRY1 plays an important role in developmental process of photomorphogenesis and light signal transduction in Arabidopsis thaliana, its function in Brassica rapa L. subsp. rapa‘Tsuda’ was not yet to be known. After blast analysis of the CRY1 gene full-length cDNA sequences of Brassica rapa L. subsp. rapa‘Tsuda’ in NCBI, a unique fragment was screened in this research. According to the Gateway clone technology, we designed two pairs of primers containing attB adapters. LA Taq DNA polymerase which can reduce the non-specific binding greatly was used during two PCR in which adding attB sequence to the cloned gene. By the BP and LR recombination reaction, the PCR product containing attB was inserted into interference expression vector pH7GWIWG2(I). The plant interference expression vector pH7GWIWG2(I)-CRY1 was successfully constructed. The results showed that it was easy to construct a plant interference expression vector by Gateway clone techno-logy. It also provided some basic information for genetic transformation with this gene and the study of its function.