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Methods for visualizing gene expression in developing animals In situ hybridization for visualization of Immunolocalization of protein expression MRNA transcripts CDNA clone Transcribe to obtain single-stranded RNA probe with modified nucleotides, Express protein of a gene in bacteria. Inject protein into complementary to MRNA sequence. vertebrate host. Extract antibodies Fixed embryos or dissected tissue (IgG) to protein. Incubate embryos with RNA probe (probe hybridizes to MRNA). Incubate embryos with antibody (antibody binds to protein). Wash away unbound probe. Wash away unbound antibody. Add enzyme- conjugated antibody to specific modified Add fluorochrome- conjugated antibody to IgG of host species. nucleotide. Wash away unbound antibody. Wash away unbound antibody. Add enzyme substrate. Visualize mRNA Visualize protein expression in fluorescence expression in light microscope. microscope. FIGURE 13-5 The two principal technologies for visualizing where a gene is transcribed or where the protein that it encodes is expressed are (left) in situ hybridization of complementary RNA probe to MRNA and (right) immunolocalization of protein expression. The procedures for each method are outlined. Expression patterns may be visualized as the product of an enzymatic reaction or of a chromogenic substrate or with fluorescently labeled compounds.

Methods for visualizing gene expression in developing animals In situ hybridization for visualization of Immunolocalization of protein expression MRNA transcripts CDNA clone Transcribe to obtain single-stranded RNA probe with modified nucleotides, Express protein of a gene in bacteria. Inject protein into complementary to MRNA sequence. vertebrate host. Extract antibodies Fixed embryos or dissected tissue (IgG) to protein. Incubate embryos with RNA probe (probe hybridizes to MRNA). Incubate embryos with antibody (antibody binds to protein). Wash away unbound probe. Wash away unbound antibody. Add enzyme- conjugated antibody to specific modified Add fluorochrome- conjugated antibody to IgG of host species. nucleotide. Wash away unbound antibody. Wash away unbound antibody. Add enzyme substrate. Visualize mRNA Visualize protein expression in fluorescence expression in light microscope. microscope. FIGURE 13-5 The two principal technologies for visualizing where a gene is transcribed or where the protein that it encodes is expressed are (left) in situ hybridization of complementary RNA probe to MRNA and (right) immunolocalization of protein expression. The procedures for each method are outlined. Expression patterns may be visualized as the product of an enzymatic reaction or of a chromogenic substrate or with fluorescently labeled compounds.

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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In Figure 13-5, two different methods are illustrated for
visualizing gene expression in developing animals.
Which method would allow one to detect where within a
cell a protein is localized?

Methods for visualizing gene expression in developing animals In situ hybridization for visualization of Immunolocalization of protein expression MRNA transcripts CDNA clone Transcribe to obtain single-stranded RNA probe with modified nucleotides, Express protein of a gene in bacteria. Inject protein into complementary to MRNA sequence. vertebrate host. Extract antibodies Fixed embryos or dissected tissue (IgG) to protein. Incubate embryos with RNA probe (probe hybridizes to MRNA). Incubate embryos with antibody (antibody binds to protein). Wash away unbound probe. Wash away unbound antibody. Add enzyme- conjugated antibody to specific modified Add fluorochrome- conjugated antibody to IgG of host species. nucleotide. Wash away unbound antibody. Wash away unbound antibody. Add enzyme substrate. Visualize mRNA Visualize protein expression in fluorescence expression in light microscope. microscope. FIGURE 13-5 The two principal technologies for visualizing where a gene is transcribed or where the protein that it encodes is expressed are (left) in situ hybridization of complementary RNA probe to MRNA and (right) immunolocalization of protein expression. The procedures for each method are outlined. Expression patterns may be visualized as the product of an enzymatic reaction or of a chromogenic substrate or with fluorescently labeled compounds.
Transcribed Image Text:Methods for visualizing gene expression in developing animals In situ hybridization for visualization of Immunolocalization of protein expression MRNA transcripts CDNA clone Transcribe to obtain single-stranded RNA probe with modified nucleotides, Express protein of a gene in bacteria. Inject protein into complementary to MRNA sequence. vertebrate host. Extract antibodies Fixed embryos or dissected tissue (IgG) to protein. Incubate embryos with RNA probe (probe hybridizes to MRNA). Incubate embryos with antibody (antibody binds to protein). Wash away unbound probe. Wash away unbound antibody. Add enzyme- conjugated antibody to specific modified Add fluorochrome- conjugated antibody to IgG of host species. nucleotide. Wash away unbound antibody. Wash away unbound antibody. Add enzyme substrate. Visualize mRNA Visualize protein expression in fluorescence expression in light microscope. microscope. FIGURE 13-5 The two principal technologies for visualizing where a gene is transcribed or where the protein that it encodes is expressed are (left) in situ hybridization of complementary RNA probe to MRNA and (right) immunolocalization of protein expression. The procedures for each method are outlined. Expression patterns may be visualized as the product of an enzymatic reaction or of a chromogenic substrate or with fluorescently labeled compounds.
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