The most compelling of these is that it is the easiest method for determining transcript size, and for identifying alternatively spliced transcripts and multigene family members. Northern analysis presents several advantages over the other techniques. Northern analysis remains the standard for detection and quantitation of mRNA levels despite the advent of more sensitive techniques. In situ hybridization is used to localize expression of a particular gene within a tissue or cell type, and RT-PCR is the most sensitive method for detecting and quantitating gene expression. (See chart.) In general, Northern analysis is the only method that provides information about transcript size, whereas NPAs are the easiest way to simultaneously examine multiple messages (Figure 1). However, the methods each have inherent advantages and/or limitations. In theory, each of these techniques can be used to detect specific RNAs and to precisely determine their expression level. Here, we review four popular methods: Northern blot analysis, nuclease protection assays (NPA), in situ hybridization, and reverse transcription-polymerase chain reaction (RT-PCR). A number of widely used procedures exist for detecting and determining the abundance of a particular mRNA in a total or poly(A) RNA sample. Zhou XL, Ruan ZR, Huang Q, Tan M, Wang ED (2013) Translational fidelity maintenance preventing Ser mis-incorporation at Thr codon in protein from eukaryote.Molecular characterization of any gene usually includes a thorough analysis of the temporal and spatial distribution of RNA expression. Yu KO, Kim SW, Han SO (2010) Engineering of glycerol utilization pathway for ethanol production by Saccharomyces cerevisiae. Weissman AM, Shabek N, Ciechanover A (2011) The predator becomes the prey: regulating the ubiquitin system by ubiquitylation and degradation. Vizeacoumar FJ, Torres-Guzman JC, Bouard D, Aitchison JD, Rachubinski RA (2004) Pex30p, Pex31p, and Pex32p form a family of peroxisomal integral membrane proteins regulating peroxisome size and number in Saccharomyces cerevisiae. Tsai PW, Yang CY, Chang HT, Lan CY (2011) Human antimicrobial peptide LL-37 inhibits adhesion of Candida albicans by interacting with yeast cell-wall carbohydrates. Tamano K, Bruno KS, Karagiosis SA, Culley DE, Deng S, Collett JR, Umemura M, Koike H, Baker SE, Machida M (2013) Increased production of fatty acids and triglycerides in Aspergillus oryzae by enhancing expressions of fatty acid synthesis-related genes. Shima S, Matsuoka H, Iwamoto T, Sakai H (1984) Antimicrobial action of epsilon-poly- l-lysine. Schmidt P, Walker J, Selway L, Stead D, Yin Z, Enjalbert B, Weig M, Brown AJP (2008) Proteomic analysis of the pH response in the fungal pathogen Candida glabrata. Özcan S, Johnston M (1999) Function and regulation of yeast hexose transporters. Kubota H, Sakaki Y, Ito T (2000) GI domain-mediated association of the eukaryotic initiation factor 2α kinase GCN2 with its activator GCN1 is required for general amino acid control in budding yeast. Khodavandi A, Harmal NS, Alizadeh F, Scully OJ, Sidik SM, Othman F, Sekawi Z, Ng KP, Chong PP (2011) Comparison between allicin and fluconazole in Candida albicans biofilm inhibition and in suppression of HWP1 gene expression. Jong AY, Chen SHM, Stins MF, Kim KS, Tuan TL, Huang SH (2003) Binding of Candida albicans enolase to plasmin (ogen) results in enhanced invasion of human brain microvascular endothelial cells. Geornaras I, Yoon Y, Belk KE, Smith GC, Sofos JN (2007) Antimicrobial activity of epsilon-polylysine against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in various food extracts. Curr Genet 40:91–109įorsberg H, Gilstring CF, Zargari A, Martinez P, Ljungdahl PO (2008) The role of the yeast plasma membrane SPS nutrient sensor in the metabolic response to extracellular amino acids. Proteomics 6:2147–2156įorsberg H, Ljungdahl PO (2001) Sensors of extracellular nutrients in Saccharomyces cerevisiae. Int J Food Microbiol 141:236–241Įbanks RO, Chisholm K, McKinnon S, Whiteway M, Pinto DM (2006) Proteomic analysis of Candida albicans yeast and hyphal cell wall and associated proteins. Chang SS, Lu WY, Park SH, Kang DH (2010) Control of foodborne pathogens on ready-to-eat roast beef slurry by epsilon-polylysine.
0 Comments
Leave a Reply. |