About Gene Cloning - EMERGENCY :(!!
Hello I have to write a report about the experiment I did aboutgene cloning and also have to answer few qns from the prof. I woudreally appreciate if anybody can help me and will thank you in apossible way. I am that desperate since this would be veryimportant for my grade :(
I don't know the insert gene and have to figure it out on NCBIBLAST search using the sequence.( I tried but I couldnt find onNCBI)
The cloning was with pGEX-4T-1 vector and unknown gene which isabout 700bp.
It says by using pGEX_3_primer, got the sequence, which would bereverse sequnece so I did reverse and it is
AAAATTAGTTTGTTTTAAAAAACGTATTGAAGCTATCCCACAAATTGATAAGTACTTGAAATCCAGCAAGTATATAGCA
TGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGTGGTGGCGACCATCCTCCAAAATCGGATCTGGTTCCGCGTGG
ATCCACCGGTCGCCACCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCA
AGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGA
GGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTC
AGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCG
AGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCT
GCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAA
GAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAG
CAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGT
GCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAA
CAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAGCGGCCGC
Then the qn is,
1.What is this gene? Where is the start and stop codon of thisgene coding sequence?
2.Would this gene translated in in-frame? Gst fusion?
3.What is the partial expected amino acid sequence of GST-YFPprotien?
4.Can you find the restriction enzyme site that we used? (BamHI,NotI)
5.To check weather the cloning was done or not, it is better touse one restriction enzyme that is on vector and the other is oninsert. In this case we used the same emzyme as we used for cloningwhich are BamHI and NotI. If we want to still use NotI but changeBamHI as different enzyme, what could it be?
About Gene Cloning - EMERGENCY :(!!
Hello I have to write a report about the experiment I did aboutgene cloning and also have to answer few qns from the prof. I woudreally appreciate if anybody can help me and will thank you in apossible way. I am that desperate since this would be veryimportant for my grade :(
I don't know the insert gene and have to figure it out on NCBIBLAST search using the sequence.( I tried but I couldnt find onNCBI)
The cloning was with pGEX-4T-1 vector and unknown gene which isabout 700bp.
It says by using pGEX_3_primer, got the sequence, which would bereverse sequnece so I did reverse and it is
AAAATTAGTTTGTTTTAAAAAACGTATTGAAGCTATCCCACAAATTGATAAGTACTTGAAATCCAGCAAGTATATAGCA
TGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGTGGTGGCGACCATCCTCCAAAATCGGATCTGGTTCCGCGTGG
ATCCACCGGTCGCCACCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCA
AGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGA
GGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTC
AGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCG
AGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCT
GCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAA
GAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAG
CAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGT
GCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAA
CAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAGCGGCCGC
Then the qn is,
1.What is this gene? Where is the start and stop codon of thisgene coding sequence?
2.Would this gene translated in in-frame? Gst fusion?
3.What is the partial expected amino acid sequence of GST-YFPprotien?
4.Can you find the restriction enzyme site that we used? (BamHI,NotI)
5.To check weather the cloning was done or not, it is better touse one restriction enzyme that is on vector and the other is oninsert. In this case we used the same emzyme as we used for cloningwhich are BamHI and NotI. If we want to still use NotI but changeBamHI as different enzyme, what could it be?
