Can anyone advise on IRES mRNA translation?

More Maixian Liu's questions See All

Can two-photon absorption/excited fluorescence happen using continuous light source?

Guys, I have a question out of my curiosity about two-photon absorption/excitation. I know that the TPA probability is dependent on the square of light intensity. Pulsed laser is usually used as...

04 May 2018 8,941 1 View

If the absorbance spectra and excitation spectra don't overlap, what info can we extract from it?

Hi Guys, I need some info regarding UV-Vis spectrum and excitation spectrum. I understand excitation spectrum is scanning the excitation wavelengths by fixing one luminescence wavelength. And...

02 March 2018 9,317 5 View

Can anyone suggest on using plvx-based lentivirus from clonetech for renilla luciferase transfection?

Hi Guys, I cloned the Renilla luciferase (hRluc from promega) into the vector of plvx from clone tech (puromycin resistance). I have tried 3 times of transfecting MDA-MB-231 cells for stably...

10 November 2016 9,649 0 View

How to separate proteins/polymers without color?

Hi Guys, I have a question regarding using Sephardex column to separate protein/ or PEI from unreacted small molecule. However, the small molecule does not have color. Is there method to tell when...

09 October 2016 1,033 2 View

How to renature proteins?

Guys, I have a question. How to re-nature the protein after the denature from GdmCl or Urea. Just curious! Thank you!

08 September 2016 9,261 8 View

Can Anyone Advise me on ompA signal peptide?

Hi guys, I am doing protein expression in BL21(DE3) and plan to use ompA signal for periplasmic protein translocation. I want to know that if it is OK to have a tag, say T7 or flag tag before ompA...

03 April 2016 5,613 3 View

Can anyone adivse me on helper phage for phage display?

Guys, I want to understand more about the function about helper phage and I start reading. One articles says that 'When wild-type phages are used, interference of the plasmid with the phage...

01 February 2016 3,949 14 View

Can anyone advise me on a dual Luciferase assay using stable cell line and lentiviral transfection?

Hi Guys, Wish someone could help me out of this. Right now I am going to have Dual luciferase transfection (Firefly and Renilla) and I want to have stable cell line that can express both of the...

01 February 2016 7,246 2 View

How to determine the diversity of phage display?

I have just entered the training of phage display and now confused with the diversity determination. As far as I know, the first round of selection has higher diversity than the second and the...

31 December 2015 4,256 9 View

I got asymmetric peak in plamonic resonance in copper sulfide nanoparticle. Why?

I have verified the crystallinity and phase purity. EDX also tells me the particle belongs to covellite with Cu:S~ 1:1. So I have no clue in the asymmetric peak of its LSPR absorbance! I also have...

06 July 2014 5,520 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Syed A Ali

Did not quite get you. IRES is used for bicistronic expression i.e., two cistrons or genes are seperated by IRES element and expressed under the control of single promoter updtream of the 1st cistron. However, I will suggest you to consider 2A peptide. Cistron downstream IRES tends to express poorly.

Maixian Liu

Thanks Ali. Maybe I should put in this way. See the patter

Cistron1----IRES----Cistron2.

Does Cistron2 need a start codon for translation into protein?

Thank you

Yes it does need its own ATG for expression.

Thanks Ali for your answer