hox基因5,UTR,3,UTR 突变如何影响多脊椎
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发布时间：2022-05-01 15:43
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时间：2022-06-18 23:13
How many bps are in the 5"UTR? It seems that you are not going to study the DNA promoter function, instead you are likely to determine translation levels of the DNA. If it is true, you need to clone your DNA containing point mutations in the 5"UTR 1) into some vectors containing T7 or SP6 RNA polymerase promoter sequences. By performing in vitro translation assays, you can examine and quantify the gene proction; 2) into some mammalian expression vector, such as pcDNA3.1 or pcDNA4. To examine the gene procts, you need to perform DNA transfection experiments using appropriate mammalian cell lines. After transfection you need to run Western blot to probe the transgene expression using antibody against the gene proct. You may need to create some tag, such as c-myc, or HA tag if your gene proct antibody is not available. If you use pcDNA4, you need to do co-transfection with another vector expressing tetracycline operator. You should add tetracycline to ince your transgene expression in the transfected cells.
