in frame cloning - cloning (Sep/16/2005 )

Hi guys,

pls help! how to check if my gene (insert) or in other words my cloning is in frame? i don't understand this in-frame thingy. i sent my sample for sequencing and got the sequence results, but how to check if it is in frame? hope someone can help me . thanks.

What are you trying to clone in frame? Your insert with a tag from the vector? N-terminal? C-terminal?
Did you translate the sequence results into protein?

this is a good explanation of what an ORF is, and should show you why it's important to have the correct ORF if you want to clone a gene and get a protein from it

http://bioweb.uwlax.edu/GenWeb/Molecular/S...ranslation.html

Wow, inframe basically means that you keeping the same codon usage for translation. If you remember the addition of amino acids to form proteins from mRNA is done from 3 base recognition codons on the mRNA, if you insert/delete one base, you will affect all downstream codons. This is what you want to prevent when you add a tag to the N-term. When you initiation translation from a tag AUG start codon, the protein you a fusing needs to be within the same coding frame as the tag. Just make sure the total number of bases in the tag is a multiple of three and you wil be in good shape.