橡胶树甲基化过滤文库的构建

doi:10.3724/SP.J.1005.2010.01071

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HEREDITAS ›› 2010, Vol. 32 ›› Issue (10): 1071-1076.doi: 10.3724/SP.J.1005.2010.01071

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Construction of a methylation filtration library in Hevea brasiliensis

HUANG Guan-Qing, HUA Yu-Wei, GAO Zhan, HUANG Hua-Sun

State Center for Rubber Breeding, Key Laboratory of Rubber Biology of Ministry of Agriculture, Rubber Research Institute of Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, China

Received:2010-01-20 Revised:2010-05-19 Online:2010-10-20 Published:2010-10-20
Contact: HUANG Hua-Sun E-mail:xjshhs@163.com

Abstract

Abstract: In order to enrich gene encoding region of Hevea brasiliensis, a methylation filtration library was constructed using Escherichia coli McrBC restriction-modification system. The titers of the non-amplified library and the amplified library were 2.6×10⁶ pfu/ml and 9.0×10⁹, respectively. The rate of positive clones was 86.4%. The lengths of inserted DNA sequence ranged from 1 kb to 2.5 kb and the average size of inserts was 1.2 kb. One hundred clones were selected randomly for sequencing, resulting in splicing out of 81 non-redundant sequences, including 6 contigs and 75 singlets. The redun-dancy was 17.35%. Blast analysis showed that 39.5% of non-redundant sequences were homologous with the Nr database, 14.81% with the EST database, and 32.1% were unknown sequences. Some sequences were related genes for flowering, insect and disease resistance. Therefore, the rubber tree methylation library is helpful for discovery and cloning of func-tional genes.

Key words: Hevea brasiliensis, methylation filtration, gene enrichment, library construction

HUANG Guan-Jing, HUA Yu-Wei, GAO Zhan, HUANG Hua-Sun. Construction of a methylation filtration library in Hevea brasiliensis[J]. HEREDITAS, 2010, 32(10): 1071-1076.

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Construction of a methylation filtration library in Hevea brasiliensis

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