Charles Goh

Vector pCambia1304 contains kanamycin resistance gene for bacterial selection, hygromycin B resistance gene for plant selection and a GUS (mgfp5:gusA fusion) reporter genes (1). From your question, you said that you want to do selection on plants. So, first, after transformation, select putative transformants from infected explants on hygromycin-containing medium plate. Resistant plants can then be used for GUS analysis.

(1) https://www.markergene.com/pcambia1304-plant-expression-vector.html

thnx for answering my question. However, i already select the transformed plant by using hygromycin as selection agent. After that, i need to select the Agrobacterium and cloning vector too. So, my question is any suggestion for both Agrobacterium and Cloning vector. Thnx.

Charles Goh What did you mean by " I need to select the Agrobacterium and cloning vector too. "? Didn't you do this before you used the Agrobacterium for transformation?

Anyway, for your binary vector selection, you use kanamycin. (see plasmid picture)

LBA4404: rifampicin and streptomycin (see below link for reference)

http://cshprotocols.cshlp.org/content/2006/7/pdb.tab194665

ok, understand dy, thnx a lot

Charles Goh A word of caution.

When you do GUS screening on putative transgenic plantlets, you need to do it a few times at different time points. For some plantlets, it will show 'positive result' (blue color) at early stage, but later on become 'negative' (no blue color). This is a 'false positive'. It is caused by some Agrobacterium that was not eradicated by antibiotics during selection. We found this during our tobacco transformation. I had to test GUS on probably first week, then second week or third week on different leaves of a 'putative' transformant to make sure they are good candidates for further Southern analysis for transgene copy numbers.

Dear Charles I think LBA 4404 has no inherent antibiotic resistance (the strain I have). After transformation with your pCambia1304 vector you can select the colonies with Kan. For the transformed plants you may use Hygromycin if your vector has resistance to it. Cefotaxime is to eradicate the bacteria from the medium and explants.