James Carrillo

11 Questions 21 Answers 0 Followers

Questions related from James Carrillo

Will agrobacterium injection / infiltration techniques yield a stable transformed line?

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 2,996 2 View

Is there a substitute for acetosyringone?

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 8,955 0 View

How long would it take for foliar application of iron to effect root gene expression?

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 2,936 4 View

How can I purify his-tagged protein from Arabidopsis?

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 4,699 1 View

Separation of small metabolites from yeast: which method to use?

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 7,612 3 View

How can I express a recombinant transmembrane protein in yeast?

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 8,394 3 View

Coupled vs. uncoupled in vitro expression methods. Why the difference?

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 9,831 2 View

How can I determine a cofactor for an inactive enzyme?

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 6,492 9 View

Are signal sequences important? (Expressing plant transporters in yeast)

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 2,882 0 View

I think Chat GPT is giving incorrect instructions...

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 6,283 3 View

Enzyme kinetics in optimized vs. nonoptimized conditions

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 7,946 6 View