Construction of a DNA library from chromosome 4 of rice(Oryza sativa)by microdissection  

作　　者：MAO YING WEI SI YUAN LIANG WEN QIN SONG XIU LAN LI RUI YANG CHEN (Biology Department, Nankai University, Tianjin 300071,China) 

出　　处：《Cell Research》1998年第4期285-293,共9页细胞研究（英文版）

摘　　要：A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification,characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method.Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000recombinant plasmid microclones from rice chromosome 4was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42%contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

关 键 词：Rice chromosome 4 DNA library MICRODISSECTION LA-PCR 

分 类 号：S511.03[农业科学—作物学] Q943[生物学—植物学]