关键词: 杉木, mRNA差异显示, 反向Northern杂交, cDNA克隆

Abstract:

Differentially displayed genes derived from the bark of two natural mutants （Cunninghamia lanceolata var. dugan and Cunninghamia lanceolata var. jurong 0）were analyzed during the wood formation by using mRNA differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Fifty-four differential display bands were collected after silver stain, and among them 29 bands were positive screening with second amplification and purification, cloning and reverse Northern blot analysis. Positive clones were sequenced and blast, the results showed: 1) nine differentially displayed transcrips matched to sequences of known or unknown function in GeneBank by Blastn (score>60). Functions of this cDNA involve ribosomal protein, signal transferring, polyubiquitin, resistant protein, histidine-containing phosphotransfer protein and ATP synthase respectively. 2) Blastx analysis showed that 12 differentially displayed fragments were able to be used for inferring functions. 3) there were still eight fragments with no hit in the GenBank. The results provided basic information for genes cloning in relation to wood forming in Chinese Fir.

Key words: Cunninghamia lanceolata, differential display reverse transcriptase polymerase chain reaction, reverse Northern blot, cDNA clone