A deep-coverage BAC library for Ponkan mandarin and its further use in screening ethylene regulated genes

作者：Wang Miao; Tan AnQun; Zhang LingYun; Yang JiaWei; Wei ZhaoXin; Cheng ChunZhen; Zhou ZhiQin; Zhong GuangYan^*

来源：Scientia Horticulturae, 2010, 127(1): 64-69.

DOI：10.1016/j.scienta.2010.09.015

• Citrus
• Bacterial artificial chromosome (BAC)
• library
• Ethylene regulated genes
• PCR screening

A deep-coverage bacterial artificial chromosome (BAC) library of Yanxiwanlu Ponkan a late maturing cultivar was constructed and evaluated Results showed that the library contained a total of 61 000 clones Restriction analysis indicated that the DNA insert sizes ranged from 30 to 170 kb with an average of 82 kb No empty clone was found Therefore the library should cover similar to 13 6-fold of the citrus genome Continuous sub-cultivation of 5 randomly chosen clones for up to 100 generations showed no detectable change in their restriction profiles To demonstrate the application of the library in discovery of genes 7 ethylene-regulated genes were screened by PCR amplification from hierarchically pooled library clones with gene-specific primers Five positive clones each representing a different gene were thus

• 出版日期2010-11-22
• 单位中国农业科学院; 西南大学