) Search online for information about HT115D(DE3) E. coli. Note that this strain
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) Search online for information about HT115D(DE3) E. coli. Note that this strain has an IPTG-inducible T7 polymerase. a.Why is the T7 polymerase necessary for dsRNA expression b.Why can we use lactose instead of IPTG to induce T7 polymerase expression? 3) Why were you advised to pick L4 or adult worms for RNAi feeding assays? netic databases: Wormbase &RNAi; feeding plasmid selection scription: There are many genetic databases publically available online where geneticists an her scientists go to share information and learn about the genes they want to study. In this la will be learning how to use Wormbase.org to identify known and likely genes in C. elegans t ant to test for a phenotype with a reverse genetics RNAi feeding assay. If you are lucky, u ay even be able to contribute to this database! ASUSExplanation / Answer
Hi
The T7 RNA polymerase promoter are extremely sensitive and specific for the genes. They have very less error rate. Hence they are desirable when inducing gene express in vitro.
The lack operon can be induced by lactose. However the lactose eventually gets catabolized to give glucose and galactose. This reduces the lac operon expression. In order to induce the operon constitutively we use non metabolizable lactose analogies IPTG. This helps in continuous lac expression.
The adult worms have remarkably simple structures. They have transparent body cavity which makes them easy tools to study anatomy. Due to simple developmental stages fate of each cell is known from embryo to larvae. Hence mutant studies using RNAi becomes extremely easy in these worms.
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