nlwxpearo
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 Posted: Sat 9:23, 15 Jan 2011 Post subject: Molecular biology techniques (Part Eight) gene exp |
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Molecular biology technology (VIII): regulation of gene expression methods
Uncle law works indicate prince Yu below to GAL4 two-phase hybrid detection system is down, a brief operation,[link widoczny dla zalogowanych], shown in Figure 4. In this system, mainly by the two yeast expression plasmid (pGBT9 and pGAD424) and a nutrient-deficient yeast strain (HF7c) composition. Wen Tian 2DN method to indicate a 1 foot prince Yu pGBT9 GAL4 of bd with tryptophan synthase gene and 3 two-phase hybridization assay gene (TRP1); pGAD424 GAL4 in the ad with a two-phase hybrid gene, also known as two-phase hybridization assay system, referred to and leucine synthase gene (LEU2); HF7c nutrition missing two-phase hybridization. It is the use of yeast cell-based gene expression system of yeast gene trap mutant strain,[link widoczny dla zalogowanych], so in the absence of leucine, the color system to detect protein interactions. We know that many acid and histidine in the medium can not grow, only when the two eukaryotic transcription factors (including yeast GALA) plasmids after co-transformation of this strain, and the reporter gene contains two in His3 spatial structure of the independent domain, expressed only after the 3 amino acids in that there is no such medium in the DNA binding domain (DNA-bindingdomain, growth, and LacZ reporter gene in this strain and both His3 bd) and transcription activation domain (transcripti0na1activa identified a gene transcription in the GAL4 under the control of regulatory sequences. tiondomain, ad). The former is to identify specific gene transcription so the two known protein genes were cloned into pGBT9 upstream regulatory sequences, and binds to the specific sequence and pGAD424, the co-transformed yeast strain HF7c, on; while the latter Other transcription factors interact, and start the base if you can not leucine, tryptophan and histidine gene transcription training. This domain 2 gene transcription start over on the growth, already showed that two protein interactions. If the process is dispensable. Using this feature. The two are to study the use of X-gal color reaction measured galactosidase enzyme activity, protein genes were cloned into two yeast expression-based plasmid can be further confirmed by protein x and protein Y of the interaction vector, and each and activators of transcription factor binding domain and activation domain with a gene linked genes in yeast co-transformation of the present, this technology is widely used in protein and protein in the mother cell to produce two fusion proteins (eg GAL4bd-X quality Interaction. If one of the plasmids and GAL4ad-Y). If the study of proteins to each other not to construct cDNA library can be screened with this technique has been used one, you can not start the yeast cell reporter gene on the genome of all known protein interactions of the protein genes. South 1, 1998 Bulletin of Biology for a 23 - Tin 4GAL4 = out of phase measurement method for the hybrid column shows the process of 4 cents prince Yu-phase hybridization assay is a three-phase hybridization assay in the two-phase hybridization assay based on the established technology , and used to study the interaction between proteins and RNA. It works with basically the same two-phase hybridization, shown in Figure 5. The difference is that the field will carry 5-phase hybrid GAL4 column method showed the work of Ha cents prince Yu Li DNA binding domain of the genes with known RNA binding protein gene (cases RevM10), known to be the RNA-binding protein The specific recognition and binding RNA genes (for example, be RevM10RNA RRERNA recognize and bind to specific binding proteins) and the RNA to study gene and the protein to study the gene and the GAL4 activation domain gene 3 gene in yeast cells expression plasmid (see Figure 6),[link widoczny dla zalogowanych], co-transformed into yeast cells and yeast cells produce two fusion proteins and a hybrid RNA. Tin for three-phase hybrid column 6 Si-law,[link widoczny dla zalogowanych], said plasmid Gan mm field with the detection process and the same two-phase hybridization with X-gal color reaction to detect the RNA molecules and to study whether the interaction of protein molecules. In addition, this method can also be used to filter a known RNA with a molecular interaction of RNA binding protein gene. 5Western blot Western blot,[link widoczny dla zalogowanych], also known as Western blot for protein analysis. It Southerrl, Northern blotting Similarities are the electrophoretic separation of the group were transferred to a solid phase support of body and amino acids are used for specific law or nuclear prince Yu 7Western printed out for the notification process to indicate the biological field 1998 '33 No. 1
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