金柑花蕾酵母双杂交cDNA文库构建及评价

为了探索金柑花发育的分子机制并研究与金柑花发育相关重要基因编码的蛋白质之间的相互作用,本研究以融安金柑不同生长时期的花蕾为材料,采用SMART技术构建了cDNA文库,对该文库进行了鉴定评价。经检测,所构建的cDNA文库滴度为2.83×10^8cfu/mL,文库库容为1.42×10^10cfu,重组率为97.91%,插入的双链cDNA片段长度主要分布在250~2 000 bp之间。结果表明,该文库达到了建库标准,理论上涵盖了全部与融安金柑花蕾发育相关的基因,为后续利用酵母双杂交技术筛选互作蛋白提供了物质基础。

Construction and Evaluation of Yeast Two Hybrid cDNA Library of Floral Buds from Rongan Kumquat

In order to explore the molecular mechanism of kumquat floral development and study the protein interaction between the important genes encoding protein relevant to kumquat floral development,the different period growth floral buds of Rongan kumquat were used as the materials in this study.The yeast two-hybrid cDNA library was constructed by SMRAT technology;then we identified and evaluated it.Based on evaluation,the titers of cDNA library was up to 2.83×10^8 cfu/mL,the number of independent clones was 1.42×10^10 cfu,the recombinant rate was 97.91%,and the length of inserted cDNA fragments ranged from 250 bp to 2 000 bp.The results showed that the constructed cDNA library met the criteria of library and contained all related genes from each period growth floral buds theoretically,and it could be used for further research of screening interaction proteins by yeast two-hybrid technology.