The invention discloses a method for constructing a
suppression subtractive hybridization (SSH)
library of
oryza rufipogon threatened by
bacterial blight germs. The method includes steps of:
sample processing, total ribonucleic acid (
RNA) extraction, mRNA purification,
suppression subtractive hybridization and
polymerase chain reaction (PCR) amplification of testing parts and driven elements, PCR products and
polarization mode dispersion 18-T (pMD18-T) carrier connection restraining, connection product conversion,
monoclonal storing and the like. The SSH
library of
oryza rufipogon threatened by
bacterial blight germs constructed by the method is high in sensitivity, even length of
insertion elements of the
library is 450bp, a large number of difference expressed sequence tags (ESTs) of the
oryza rufipogon threatened by the
bacterial blight germs can be obtained fast in one step, partial sequence information is provided for separating overall-length cDNA sequence of clone bacterial
blight resisting related genes, simultaneously important theoretical basis for improvement of
biology is provided and new bacterial
blight resisting related genes can be obtained.