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Questions related from James Carrillo
I've recently come across the method of agroinjection and vacuum infiltration of various plant tissues (seedlings, potato eyes, tomato fruit). These methods are new to me and very fascinating but...
07 May 2024 3,108 2 View
I could've sworn that I read a protocol where strained plant "juices" were added to agrobacterium cultures in lieu of pure acetosyringone. If you have heard of anything like this, please let me...
07 May 2024 9,081 0 View
I'm thinking 2 hours. This is my first qPCR experiment on plants and I'm hoping to catch the earliest changes in root gene expression when iron is applied to leaf tissue. I've found a similar...
19 July 2023 3,023 4 View
I see that some pCAMBIA plant expression vectors feature histidine-tags. I am having a difficult time finding examples of purification of tagged proteins from plant tissue. I have read examples...
24 May 2023 4,760 1 View
I'm trying to separate small metabolites from yeast lysate prior to FMOC-derivatization and HPLC analysis. The cleaner I can get the lysate the better but I'm on the fence when it comes to buying...
22 May 2023 7,672 3 View
I am modifying the yeast expression vector pYES2 to express a plant transporter. The gene is cloned but I failed to insert an appropriate signal sequence. If the recombinant protein contains...
25 January 2023 8,448 3 View
If you have perform an in vitro transcription reaction, using a T7-RNA polymerase, the buffer/solution doesn't appear to contain any inhibitors or issues for translation. Why then must then the...
22 November 2022 9,893 2 View
Intro SAM-synthetase requires two substrates, L-methionine and ATP, to form S-adenosylmethionine. All other information aside, IT REQUIRES TWO SUBSTRATES! So determining Km for each substrate...
20 June 2021 291 1 View
I am having issues restoring activity to an extradiol dioxygenase that I am studying. It is a recombinant form of a Rhizobium enzyme, expressed with a N-terminal 6xHistag in E. coli BL21 cells....
01 February 2016 6,557 9 View
In heterologous expression studies, I am reading through examples of plant transporters which are produced in yeast. I was under the impression that proteins destined for the cell membrane...
01 January 1970 2,929 0 View
I tried using ChatGPT for the first time...thought maybe it could provide something different than what I planned. I think it is giving really wrong information. Wouldn't using acetone precipitate...
01 January 1970 6,329 3 View
I work with a thermophilic enzyme, its optimal temperature is 50 C. From some assays at 50 C and at 35 C, I was able to construct two lineweaver burke plots. I want to ask you if my...
01 January 1970 8,015 6 View
