BIOL 3010 Lecture Notes - Hypoxanthine-Guanine Phosphoribosyltransferase, Blastocyst, Thymidine
29 Jan 2013
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In-situ hybridization (rna) cross link proteins and remove lipids to allow access use radioactivity, fluorescence or colorimetric assays can be used to figure out timing of gene expression. Immunohjistochemistry (protein) location and timing of protein expression. Marker/reporter genes product must be low background (specific) and low impact (non-toxic) lacz (product is blue) Gfp can be toxic, but also expressed as fusion-proteins. Requires that promoter for normal gene expression is known. Ligate reporter gene to the upstream controlling elements. Introduce marker gene/promoter to all cells (i. e. transgenesis) Can use coat color, eye color, lacz, gfp, etc. to identify presence of chimericism. Each tissue type is unique as to the extent of intermingling. Es technology (embyronic stem cells) derived from the icm and can proliferate undifferentiated in cell culture can be manipulated and may contribute to germ cells in chimeric models usually are male embryos that survive.
