which part of gene is needed for normal expression in vector - (May/22/2008 )
Hi,
I want to express sugar transprot gene under the control of Hsp promoter vector in plant.i want to see the expression of this under high temperature. which part of gene i needed to clone into vector contaning hsp promoter my gene is f 1426 bp length. how can i design primers to isolate that part. my final aim is to increase sugar availability in plants.please help me in this issue and thank you very much for your replies.
SVRREDDY
Ph.D Student
I want to express sugar transprot gene under the control of Hsp promoter vector in plant.i want to see the expression of this under high temperature. which part of gene i needed to clone into vector contaning hsp promoter my gene is f 1426 bp length. how can i design primers to isolate that part. my final aim is to increase sugar availability in plants.please help me in this issue and thank you very much for your replies.
SVRREDDY
Ph.D Student
Generally, cloning the Open reading frame (ORF - start codon to stop codon) under a promoter is sufficienct. Most vectors provide promoter (hsp here) and a poly-adenylation signal. You may also decide to clone the entire gene (including 5' and 3' non-coding regions, not the introns). Sometimes it helps in achieving higher expression, higher stability.
Hi cell counter,
Thank you very much for your reply regarding my question,as like your reply i found orf regione in my gene it is of 1116 bp size.I designed primers to isolate that with PCR but the problem is with primers is primer product size is only 170 to 190bp, if it is like this how can i isolate this fragment.please help in this issue and give your suggestions in this.
Thanking you very much.
SVRREDDY
Ph.D Student