用cDNA-AFLP及其改进的方法分析茶树花发育过程中的基因表达

摘要

A comprehensive transcriptome analysis by means of cDNA-amplified fragment length polymorphism (AFLP) and modified cDNA-AFLP technique were performed in order to gain further understanding of the development in molecular level of tea reproductive organs: tea floral bud. Fragments detected among the young and the mature tea flo-ral bud were grouped into three differential expression patterns : (Ⅰ) The young-specific expressed fragments; (Ⅱ) Fragments detected only in the mature tea floral bud; (Ⅲ) Fragments occurring in both. Some transcript-derived frag-ments (TDFs) were isolated by cDNA-AFLP and its modified technique. The majority of the TDFs are still function un-known according to BLAST alignment search in public data bases. Some of them showed homology with known genes. They are anthranilate pbosphoribosyl transferase (AnPRT), beta-primeverosidase, Cytchreme (P450), DnaJ-like pro-tein, Ribulose-1,5-bispbosphate carbexylase/oxygenase small subunit (RbcS), alpha-tubulin and Carbonic anhydrase by cDNA-AFLP technique. We isolated pollen coat protein gene(Pcp) by modified eDNA-AFLP method. The full length of three genes, pollen coat protein (Pcp), DnaJ-like protein and RbcS, have been cloned and submitted to Gen Bank.%利用cDNA-AFLP及其改进的cDNA-AFLP方法,分析茶树花发育过程中的基因表达.其发育过程中的基因表达可以分为3类:未成熟阶段发育特异基因;成熟阶段发育特异基因;茶树花发育过程中均表达的基因.利用改进的cDNA-AFLP方法,我们获得编码花药发育特异基因:pollen coat protein(Pcp).用cDNA-AFLP方法,我们获得7个已知功能基因分别编码Cytchrome(P450),beta-primeverosidase,Dnaj-like pro-tein,anthranilate phosphoribosyi tramferase(AnPRT),Ribulose-1,5-bisphosphate carboxylase/oxygenasesmall subunit(RubpS),alpha-tubulin和Carbonic anhydrase.用RACE方法获得pollen coat protein(Pcp),DnaJ-like protein和Ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit等3个基因的全长,并已提交GenBank.