Pat Brown developed a technique where cDNAs can be embedded onto lass slided.Using these DNA arrays,he could do large-scale expression studies
div style="text-align: justify;">Growth stage-specific mRNA are isoloated,and then reverse transcribed to give inique cDNA populations.These are directly embedded onto specially cated gass slides. Dna Array
These slides are coated with poly-lysine,which is positively charged.DNA is negatively charged,so that cDNA "sticks" to the slide through an ionic interaction.The cDNA can still interact with DNA probe.
Brown used srrays to obtain gene expression profiles of cancer.Diffuse large B-cell lymphoma(DLBCL) is a common lymphoma-cancer of the lymp nodes.He found sub-types of DLBCL that correlated with survival rates.
Brown made a chip with genes expressed by the lymph nodes and those importtant in cancer biology .a total of 17856 cDNA genes were printed onto what we called "lymphochip".Remember ,each square on teh chip corresponds to a differnt cDNA.
Then ,he made CDNAs from different DLBCL tumors.
He labelled one set of cDNA with a red florescent tag;the other with a green tag.
Brown incubated the arrays with the tagged cDNAs, which bound to the matching genes printed on the array.
Since he knew the positions of the genes on the DNA array,Brown could figure out the levels of gene expression based on the color signal.If the gene was only expressed in DLBCL1 cells,the square was red.similarly ,of the gene was only expressed in DLBCL2 cells,the square was green.If the gene was expressed equally in both cells,the square was yellow
Thus ,brown identified two sub-types of DLBCL-GC B-like and Activated B-like DLBCL.These sub-types have different responses to the therapy,and with this type of diagnosis,more tailored treatment can begin for patients.This type of tailored treatment is called "pharmacogenomics."
Brown can also analyse the differences in gene expression for these two very similar lymphomas.This may give better understanding of how cancers work, and hopeflly develop beter therapies and cures.
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