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- W2381771439 abstract "To investigate the role of TGF β1 in hematopoitic regulation,TGF β1 partial gene fragment were cloned into T_vector,~(32)P_labled TGF β1 partial transcripts as target RNA was Transcripted in vitro and purified by denaturing polyacrylamide gel electrophoresis(PAGE).Anti_TGFβ1 ribozyme were designed by software pc_FOLD combined with the data from NCBI rat GenBank.Thereafter,synthetic ribozyme fragment was cloned into pGEM_9Zf(-) at downstream of T7 promoter,to asses the cleavage activity of TGFβ1,ribozyme fragment was PAGE_purified and incubated with target TGFβ1 fragment with equimolar amount at different conditions.Bands on denaturing PAGE corresponding to substrate and cleavage products were analyzed by autoradiogrphy.Wide_type ribozyme(Rz445) completely cleavaged TGFβ1 mRNA while single base mismatch(Rz445m) significantly reduced the cleavage efficiency.Further examine found that Rz455 possessed cleavage activity at 37℃ with K_m 29.55 nmol/L and K_(cat) 0.1533 min~(-1),respectively.The optimal temperature was at 50℃.The cleavage efficiency was enhanced as the incubation period increased within 40 min(CE_(max)=32.67%).These results indicated that ribozyme might suppress the expression of TGFβ1 in vivo,it might be developed as a potential candidate for the stem cell transplantation." @default.
- W2381771439 created "2016-06-24" @default.
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- W2381771439 date "2006-01-01" @default.
- W2381771439 modified "2023-09-23" @default.
- W2381771439 title "Cleavage Activity Analysis of Hammerhead Ribozyme Targeted to TGF β1 mRNA in vitro" @default.
- W2381771439 hasPublicationYear "2006" @default.
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