Please come prepared...bring all the questions you have about BIOL335!
* operons and bacterial gene regulation
* restriction mapping
* cloning (general approaches)
* techniques (Southers, northerns, transformations, selections, PCR, RT-PCR, footprinting, colony lift hyb., indicator-based assay systems, chromosome walking...)
* eukaryotic gene regulation and development
* genomics (molecular markers, contigs, sequencing of whole genomes...)
* GPCRs
Happy studying!
13 comments:
Hi Pam,
I'm worried that we are not getting an exam package from the biosoc (or so is the case at the moment)... but do you think the package won't really help us anyways because of the different materials covered in regular sessions and summer sessions?
Some of the questions won't apply to you, because the course was a bit different. I think you should be fine even without the biosoc package!
do we need to know about the yeast-2-hybrid system of euk. regulation?
do we need to know about 5' UTRs of biocoid, nanos?
thanks
Hi Pam,
I am wondering when we are cloning with P element, we make the one of the constructs with IR, promoter of the gene of interest, Reporter Gene, and Marker gene right? but I'm confused because I was looking at the lecture notes from spring, and it doesn't mention marker gene. It says that the Reporter gene is what we will be screening for in the colonies, such as for LacZ. So my question is...if we are using a Marker gene, then what is the purpose of the reporter gene?
Thank you!
You don't need to know about the yeast 2hybrid system (nobody uses it anymore, anyway), but you'll need to know the principle about the UTRs of bcd and nanos. You need to keep in mind that you can have specific proteins binding to an RNA's UTR and 'taking it' to a specific location in the cell.
The reporter gene vs. marker gene:
sometimes people call the marker gene a 'reporter' (to add a little bit of confusion), but if we call things like we've done in class, you are corrrect, you always need a marker, but don't always need a reporter (only need it for specific experiments).
Also, sometimes people give it for granted that you'll have a marker gene in the modified P element. A bit like people assume that if you are using a plasmid, OF COURSE you'll have an antibiotic resistance gene in it (to let you select for the bacteria that received the plasmid).
Cheers
Hi Pam,
I want to ask you about VNTR markers in comparison to RFLP markers. For VNTR, when 2 alleles co-segregate together, it means those 2 alleles must be from different genes.
In contrast, for RFLP markers, if 2alleles cosegregate together, it means they are from the same gene.
Is this correct?
Thank you Pam!
I assume that by 'alleles' you mean 'bands on a gel'...
If this is the case, yes, sort of. If it is possible to inherit 2 particular VNTR bands from the same parent, then they are not allelic to each other, meaning they are at 2 different locations on the chromosome (like 2 genes).
If you inherit 2 RFLP bands from the same parent, then these 2 bands represent one RFLP locus (i.e. a region that hybridizes to the probe).
I hope this helps-and hopefully I interpreted your question correctly.
Cheers
Pam
Hi
If we want to use P element to clone the leg gene of Drosophila, and we use the red eye gene as the marker do we need to include the enhancer for the red eye gene on the modified P element? Where does thee transcription factor for the red eye gene come from? How does the leg enhancer regulate the eye gene on the P element?
Yes, you'll need to include at least some enhancer sequences for the marker gene (but we always use genes that we know very well as maarkers....so we know their enhancers too, and it's not too hard to do).
The TFs are going to be in the cells that will make the eye, the fly makes all the right TFs in the right cells.
There is no leg enhancer in the P element...but I guess you mean if the P lands in a legs specific gene, then its enhancer will be able to interact with the promoter of the marker...right? Well, it may make the marker expressed in the legs too, but in most cases we won't see anything (all the other genes necessary to make, for example, red tissue, are not active in the legs).
EXCELLENT QUESTION, BY THE WAY!
When we are asked to clone a gene, do we need to include the regulatory region (ex. enhancer) or just the coding region?
Thanks
Good question. if nothing else is specified, you'll need to clone the gene with its promoter.
You're welcome!
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