Somethings about P-element (Urgent, Thanks@@) - (Feb/19/2008 )
In order to engineer Drosophila with P-element:
In the transformation vector, there are 5' and 3' P-element terminal inverted repeats, I want to know what is the function of terminal inverted repeats?
2. Secondly. I want to know the principle of this technique. Since both transformation vector and helper plasmid (Transposase source) are injected into embyro if Drosophila, I want to know where has the cloned DNA (in vector) gone? Does the cloned DNA jump over all the genome in the embyro
3. In the P-element vector, there is Reporter gene (e.g LacZ), acutally what is the function os this transgenic reporter gene? In my teaching notes, it said it can be used to detect immune-induced genes, I would like to know the principle.THanks
I want to know what is the function of terminal inverted repeats?
the inverted repeat sequence is at the beginning and the end of every transposon. this special sequence is recognized by the transposase enzyme so it copies the right area in the genome (or p-element in your case).
2. Secondly. I want to know the principle of this technique. Since both transformation vector and helper plasmid (Transposase source) are injected into embyro if Drosophila, I want to know where has the cloned DNA (in vector) gone? Does the cloned DNA jump over all the genome in the embyro
the gene of interest is inserted randomly anywhere in the genome. correct me if im wrong! i never did this stuff with drosophila but similar methods with other cells allways result in a randomly located insert.
3. In the P-element vector, there is Reporter gene (e.g LacZ), acutally what is the function os this transgenic reporter gene? In my teaching notes, it said it can be used to detect immune-induced genes, I would like to know the principle.THanks
the LacZ gene is coding beta-galaktosidase. you should know what this enzyme is for. its quite basic knowledge. there are some assays using lacZ as a reporter. google the word "x-gal" if you want to know more.
and dont be confused. the lacZ is for the E.coli that you need to multiply your helper plasmid. drosophila probably cant even express this gene!
hope i was helpful
the inverted repeat sequence is at the beginning and the end of every transposon. this special sequence is recognized by the transposase enzyme so it copies the right area in the genome (or p-element in your case).
2. Secondly. I want to know the principle of this technique. Since both transformation vector and helper plasmid (Transposase source) are injected into embyro if Drosophila, I want to know where has the cloned DNA (in vector) gone? Does the cloned DNA jump over all the genome in the embyro
the gene of interest is inserted randomly anywhere in the genome. correct me if im wrong! i never did this stuff with drosophila but similar methods with other cells allways result in a randomly located insert.
3. In the P-element vector, there is Reporter gene (e.g LacZ), acutally what is the function os this transgenic reporter gene? In my teaching notes, it said it can be used to detect immune-induced genes, I would like to know the principle.THanks
the LacZ gene is coding beta-galaktosidase. you should know what this enzyme is for. its quite basic knowledge. there are some assays using lacZ as a reporter. google the word "x-gal" if you want to know more.
and dont be confused. the lacZ is for the E.coli that you need to multiply your helper plasmid. drosophila probably cant even express this gene!
hope i was helpful
It is very useful!!!!!THank very much!!