Isolation Of Differentially Expressed Genes Of Straw Mushroom In Low Temperature Condition By Modified CDNA-AFLP Method And Isolation Of Ras Promoter Of Straw Mushroom

Using a single restriction enzyme based on cONA-AFLP to isolate the differential gene, we got 13 differentially expressed gene fragments, we named them, Vl(488bp), V2(551bp), V3(557bp), V4(285bp), V5(173bp), V6(604bp), V7(743bp), V8(836bp), V9(709bp), V10(about720bp), V11(1077bp), V12(l077bp), V13(617bp). Vll and V12 are the same gene fragments. Using BLASTn to analyse these sequences, we got the following results: VI, V2, V3, V4, V5, V6, V7, V9, V10, 11(V12), V13 are the unknown gene. V8 is 84-93% the same as ATPase gene in a region of 312bp. Using BLASTx, we know that: V1, V2, V3, V4, V11(V12), V13 are more than 30% the same as the protein of other fungus. V3 is about 33% the same as cytochrome P450(from Coprinopsis cinerea ), V8 is about 68% the same as ca2+-ATPase (from Neurospora crassa).Using PCR technology to isolate ras promoter, we got a DMA sequence that is likely to be a promoter sequence. This DNA sequence is not the same as other ras promoter sequences in edible mushroom. Two possible foundational promoter sequences locate on the region of 243-293bp and 539-589bp.There are two transcription start points. One locate at the site of 283bp,another locate at the site of 579bp. There are also many important cis-acting regulatory elements, such as TATA-box, CMT-box, G-box, ABRE, WUN-motif, HSE, P-box, TATC-box, I-box et al.