how can i construct double mutant or triple mutant in rice

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
elizebeth
Garter
Garter
Posts: 2
Joined: Sun Jul 23, 2006 7:15 am

how can i construct double mutant or triple mutant in rice

Post by elizebeth » Sun Jul 23, 2006 7:23 am

I want to obtain double or triple mutant in rice. The genes are highly homologous. I want to use RNAi method. But i didn't know whether the effect is obvious or not. I expect someone can give directions to me.

User avatar
LilKim
Coral
Coral
Posts: 436
Joined: Mon Mar 20, 2006 8:36 pm
Contact:

Post by LilKim » Mon Jul 24, 2006 4:22 am

I don't know much about plants but you'd have to stably transfect a vector (introducing a transgene) that stably expresses miRNA homologous to the 3'UTR of your target genes.

i've never done this before, so i think you'd have to look for programs (or a reference) that would help you to design a vector that would result in the correct miRNA for you project. (cuz it isn't exactly straight-forward ... although, I don't remember it being difficult fromt he literature that I have read)

A good general "starting" reference for miRNA:
http://ambion.com/main/explorations/mirna.html

i also read an EXCELLENT paper on designing miRNA vectors. (Please let me know if you'd like me to 'hunt it down' ... it's tucked away in one of my HUGE binders somewhere... so i'd have to take a moment to look for it, if you need me to)

good luck and let me know?!
- KIM

elizebeth
Garter
Garter
Posts: 2
Joined: Sun Jul 23, 2006 7:15 am

Thank you!

Post by elizebeth » Mon Jul 24, 2006 11:46 am

I had read a paper about amiRNA. It's name is
"Highly Specific Gene Silencing by Artificial MicroRNAs in Arabidopsis"
But this method can only applied to Arabidopsis now. I don't know some better way so far. You need not find that paper for me. It is very kind of you to give some advice to me. I am not familiar with miRNA. May I should read sth about it. Thank you! :D

User avatar
LilKim
Coral
Coral
Posts: 436
Joined: Mon Mar 20, 2006 8:36 pm
Contact:

Post by LilKim » Mon Jul 24, 2006 7:48 pm

So, I know nothing about plant biology/genetics and I'm curious... why can't the arabadopsis miRNA knock-down technique work in rice?

qq-wheat
Garter
Garter
Posts: 7
Joined: Fri Jul 14, 2006 5:20 am

Post by qq-wheat » Wed Aug 16, 2006 2:09 am

I know something about transgene ,but i don't konw something about miRNA. May be you can get some information from magazine,such as scinece.

Post Reply

Who is online

Users browsing this forum: No registered users and 7 guests