Lecture 8 - PLB 113 - Lecture 8 II. Gene Transfer and...

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PLB 113 - Lecture 8 II. Gene Transfer and Epigenetic B. Agrobacterium tumefaciens Biology C. Development of a Gene Transfer System D. Transgene Expression - Soybean Lectin -------------------------------------------------------------------------------------------- Tobacco mDNA and null mDNA experiment: = 60000 genes were expressed = represent 12% scDNA = may not represent all the genes in Tobacco = this suggests that a lot of scDNA doesn't correspond to the genes DNA microarray: - measure both qualitative and quantitative aspects of gene expression 1) take mRNA from particular tissue or organ 2) make cDNA from mRNA and label the cDNA 3) take the chip that has the representation of the entire genome 4) the complementary cDNA will hybridize with the complementary DNA on the chip 5) the hybridized spot will light up. Intensity of spot => how much gene you have. Since we know what probe is present on which spot of the chip => know what genes are expressed Most of the mRNA are in common between all the plants Common mRNA are qualitatively regulated => determines functions and morphology of the organs. * primarily expressed in a given order of the plants Next-generation sequencing: - take mRNA and make ds cDNA from it - break cDNA - sequences = have lots of read - identify the presence of mRNA Question of the Day: Do Next-generation RNA sequencing experiments provide information about the level of each mRNA in a population? Explain your answer. YES. Adaptors stick on the end of mRNA = tell the beginning and end of mRNA = tell how many represent a single copy of mRNA vs. many pieces of mRNA. high level of mRNA => lots of reads low level of mRNA => less reads
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Because cDNA generated should be representing mRNA. More mRNA = more cDNA = more sequences (reads) -----------------------------------------------------------------------------------------------------------(MT1- END) B. Agrobacterium tumefaciens Biology A. tumefacian infect wounded plant => upon infection, the tumor (crown gull) is induced on the plant = T-DNA from bacteria has been integrated into plant genome. Overview of infection: - wounded plant release singles - singles attract the Agrobacterium to the wound site and also induce the gene in the Agrobacterium to begin the infection process - bacteria attach the plant cell - T-DNA move from bacteria into the plant cell (plant genome) - integration of T-DNA into plant genome results the growing of crown gull. Ti Plasmid
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Lecture 8 - PLB 113 - Lecture 8 II. Gene Transfer and...

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