基因表达的分析中的内参照设置.pptVIP

基因表达分析中的内参照设置 内参照基因的作用 内参照基因的条件 成功的实验设计要求的对照实验 阳性模板对照 阴性对照 阳性基因对照 内参照 阳性基因对照的阴性对照 内参照基因的概念和特点 所谓内参照,就是实验材料本身固有的,在实验的全程与目的基因相伴随,接受同样的处理,并可作为参照的基因. 特点1:在材料受到不同的(实验处理因素)过程,表达恒定, 特点2:在动物的各种组织中表达比较恒定, 特点3:除实验设计处理因素外,能够与目的基因一起受到同等程度的非设计其他因素影响 外参照基因,人工加入的质控成分 不管是何时加入的都是外参 好的内参的额外要求 表达丰度与目的基因接近 在各个物种中序列较为保守,突变机会小 功能重要 最好是管家基因 能够在多种检测中通用 操作方便,结果漂亮,重复性好 常用内参基因 ACTB (actin, beta) GAPDH 16Sr 18Sr TFRC(CD71) TBP(TATA-BOX BINDING PROTEIN) B2M(BETA 2 MICROGLOBULIN) PGK1(phosphoglycerate kinase 1 ) GAPDH Glyceraldehyde-3-phosphate dehydrogenase: catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The enzyme exists as a tetramer of identical chains. A GAPD pseudogene has been mapped to Xp21-p11 and 15 GAPD-like loci have been identified. B2M Beta-2-microglobulin is a serum protein found in association with the major histocompatibility complex (MHC) class I heavy chain on the surface of nearly all nucleated cells (Gussow et al.,1987 [PubMed 3312414]).[supplied by OMIM] TFRC（CD71）TRANSFERRIN RECEPTOR Function: Cellular uptake of iron occurs via receptor-mediated endocytosis of ligand-occupied transferrin receptor into specialized endosomes. Endosomal acidification leads to iron release.The apotransferrin-receptor complex is then recycled to the cell surface with a return to neutralpH and the concomitant loss of affinity of apotransferrin for its receptor. Transferrin receptoris necessary for development of erythrocytes and the nervous system (By similarity). A secondligand, the heditary hemochromatosis protein HFE, competes for binding with transferrin for anoverlapping C-terminal binding site MT-RNR2 mitochondrially encoded 16S RNA 18S ribosomal RNA LOC100008588 gene The sequences coding for ribosomal RNAs are present as rDNA repeating units in the p12 region of chromsomes 13, 14, 15, 21 and