REGULAR ARTICLES
Cell Research (1998)8:258-293
© 1998 SIBS, CAS All rights reserved 1001-0602/98
Construction of a DNA library from chromosome 4 of rice (Oryza
sativa) by microdissection.
Mao YW, Liang SY, Song WQ, Li XL, Chen RY.
Biology Department, Nankai University, Tianjin, China.
Correspondence:
Chen RY
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Chen RY
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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 100,000 recombinant plasmid microclones from rice chromosome 4 was
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.
Keywords : Rice
chromosome 4, DNA library, microdissection, LA-PCR. |
