I have isolated salinity related gene from halophyte and I have cloned them in pAUR123 (protein expression vector). Now I want to introduce them into yeast (Saccharomyces cerevisiae) to study the sodium uptake experiment and yeast growth experiments under salinity stress.
Any one know which yeast strain should be used for this study?
Please kindly guide me.
Thank you !
1. Do you know what is the possible 'protein class' of your salinity-related gene product?
2. Attached paper describes that the transformants of yeast strain W303.16 with a Arabidopsis cDNA increase the Na+ efflux, and showed a modest but clear increase of NaCl tolerance (from 300 to 500 mM NaCl in arginine phosphate [AP] medium) [see attachment].
Can you transform this yeast with your pAUR123 to see any Na+ efflux increase?
Hi Yuan. Thank you for your answer. Do you know how to buy this strain W303.16?
After searching few article i came to know that Saccharomyces cerevisiae strain 9.3 (ATCC, USA, No. 201409) is best to study these kind of experiments. But I don't know how to buy that strain. In online ATCC database it is showing it is unavailable:-(
Hi Harshraj,
When I used ' Saccharomyces cerevisiae strain 9.3 ' to search ATCC database, this one jumped out (see below), do you know that they are the same thing? They might have changed their category #.
Saccharomyces cerevisiae strain 9.3 (1 matches)
YCpMATa (ATCC® 77172™)
http://www.atcc.org/Search_Results.aspx?dsNav=Ntk:PrimarySearch%7cSaccharomyces+cerevisiae+strain+9.3%7c3%7c,Ny:True,Ro:0,N:1000552&searchTerms=Saccharomyces+cerevisiae+strain+9.3&redir=1
1. This group also used Saccharomyces cerevisiae strain 9.3 as their experimental material (a NaCl sensitive strain) for salinity tolerance research. Attached is their 2014 paper. If you cannot buy this strain, may be you can ask this strain from them directly.
2. I am not sure where you can buy W303.16 yeast strain. I searched on ATCC, they did have something called 'W303', but without '.16'. So, I am not sure they are the same, or with different genotypes.
Thank you So much Yuan for all your efforts.
ATCC® 77172™ is not Saccharomyces cerevisiae strain 9.3 that is another mutant. Unfortunately ATCC is no more producing Saccharomyces cerevisiae strain 9.3 from 2013. :-(
Well I have asked to this spain group of 2014 article I am just waiting to get their answer.
Hi Harshraj,
1. Also look at the first attached paper. The authors have screened full-length cDNAs from halophyte plant P. tenuiflora in yeast. Two cDNA were chosen and cloned into yeast-expressing vector [ pYES2-NaCl-158 and pYES2-NaCl-167 ], and transferred into yeast strain INVSc1 for salt-tolerant test. The results of testing on NaCl look pretty good (see Fig. 6).
2. This yeast strain INVSc1 can be purchased at ThermoFisher:[ https://www.thermofisher.com/order/catalog/product/C81000 ]
3. I also attach a second paper (2015), where the research project is pretty much the same as the first paper listed above. The only difference is that they expressed the candidate cDNA in plant Arabidopsis.
4. Is your pAUR123, a yeast-expressing vector? I saw both paper used pYES as yeast-expressing backbone.
5. After testing in the yeast, are you going to test your cDNA in plant, such as Arabidopsis?
Thank you once again Dr. Yuan fro all your efforts.
1) pAUR123 and PYES2 are almost same. pAUR123 is kind of old yeast expression vector.
2) Yes I have cloned my gene into plant expression vector also to express it in Arabidopsis.
I will try to order the yeast strain INVSc1
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts