mRNA差异显示法研究杉木木材形成相关cDNA

Isolation cDNA Related to Wood Forming in Chinese Fir (Cunninghamia lanceolata) by mRNA Differential Display

利用差异显示法(DDRT-PCR)研究杉木的2个自然变异类型(句容0号及独干杉)木材形成过程中基因表达差异。通过银染后切割回收54个差异条带,经2次扩增和纯化、克隆转化及反向Northern杂交验证后共获得29个阳性克隆。测序后进行比对分析,结果表明:1)Blastn比对(分值>60)有9个cDNA克隆在GeneBank中找到了相似的功能,分别与核糖体蛋白基因、信号传导功能、泛素蛋白基因、抗性功能、组氨酸磷酸转移蛋白、细胞发生功能及能量代谢相关;2)Blastx比对有12个序列可进行功能推测;3)还有8个cDNA在数据库中未发现匹配信息。这些信息可为杉木木材形成相关基因的分离和克隆提供研究基础。

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.