Questions tagged [molecular-biology]
The study of the molecular processes underlying life.
1,605
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Is it possible to use two forward primer and one reverse primer?
I have eGFP and the gene fragment i assembled by pcr.The next step is that i have to join my fragment fragment into 3'p end of the eGFP.I have three primers FP1 is forward primer for eGFP, FP2 has ...
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0answers
23 views
Is sickle cell haemoglobin more insoluble than normal haemoglobin?
I learnt in school that haemoglobin is soluble due to the hydrophilic R-groups pointing outwards and for folks with sickle cell anaemia the there's a mutation that causes there to be a change in the ...
7
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1answer
358 views
Why do OX5034 GM mosquitos require the presence of tetracycline to survive? What does the drug do in this case?
I'm confused. Debug Fresno; why are the released mosquitos said to be sterile? from 2017 addresses male mosquitos released with a bacteria that will affect fertility of females after mating. They are ...
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0answers
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See image to answer question [closed]
Can you please help me understand this question I have! Thanks!
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1answer
37 views
Can a constitutively active kinase be highly regulated?
I am studying the protein kinase GSK3 and I am learning about the regulation of its activity. Many journal papers that I have read have stated that GSK3 is unique because it is a constitutively active ...
2
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1answer
34 views
Can oxytocin travel from one cell to another via gap junctions?
Oxytocin is a 9-residue secreted peptide. As a hormone, can it travel through gap junctions, assuming that it is stored in pre-synaptic neuronal vesicles?
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28 views
Is there “stem” organelles in plant meristems?
Two cytoplasmic genomes exist in plant in addition to the nuclear one. As far as I understand, they divide more or less during the entire life of the plant along with cell proliferation/growth in ...
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0answers
26 views
primer sequences for detection of mutations [closed]
I am very new to this world of molecular biology. I wish to search for a primer sequence for detecting KRAS and EGFR mutations. Kindly let me know how can i do that?
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1answer
40 views
Can GFP reporting be used to track localization of peptides in the ER, Golgi, and plasma membrane?
Suppose I want to study the trafficking of a peptide throughout the ER, Golgi, and plasma membrane. An idea I had was labeling a secreted or plasma membrane integral protein with GFP and using time-...
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0answers
17 views
How does flow rate of Ion channels depend on the length of the selectivity filter?
The length of the selectivity filter of ion channels such as the potassium channel commonly is only half of the thickness of the membrane. Some book such as MBoC says this can benefit the transfer, so ...
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64 views
How can I clone a gene into a plasmid vector with an N-terminal his tag and TEV cleavage site between the tag and the start of the sequence?
I'm a scientist who has significant experience in chemistry but am relatively new to molecular biology and biochemical techniques. I'm trying to make an isolated domain of a protein (166 residues, 19....
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1answer
77 views
Why is DNA replication not 100% accurate
I've been reading about DNA mismatch repair (MMR) and how this process improves DNA fidelity. However, I was wondering, what is stopping MMR from correcting all mistakes in the daughter DNA with 100% ...
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3answers
377 views
Does the term 'protein expression' refer to the production of proteins only or also their regulation?
I am learning about molecular biology and I have come across the term 'protein expression' in a research paper. I have searched the definition of this term online and on the Thermo Fisher Scientific ...
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isn't GST affinity to GSH only temporary in pull down assay?
I'd expect that as all enzymes, GST would release GSH shortly after binding. Or does it release it only after 3rd component is met to which GST transfers GSH and that component is missing in GST pull ...
0
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1answer
28 views
What are the uses of centrioles and matrix of centrosomes?
In centrosomes, the gamma-tubulin ring complexes (gamma-TuRC) located on the surface are essential as the nucleation site for the growing (with polarity) of microtubules.
However, I see that there is ...
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0answers
21 views
How do cilia beat?
From what I have learnt, the mechanism of the beating of cilia is: movement of dynein towards the (-)-end i.e. towards the microtubule-organizing center (MTOC), causing the 2 pairs of microtubules to (...
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0answers
21 views
When transmembrane proteins destined for the plasma membrane are in the ER membrane are they in their final folded form?
I am studying how transmembrane proteins are made and I have read that proteins that are destined for the plasma membrane are initially in the ER membrane and do not get translocated into the ER lumen ...
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1answer
30 views
What is meant by 'topologically continuous' in the context of cell compartmentalisation? [closed]
I am reading the textbook 'Molecular Biology of the Cell', and I am reading the chapter about the compartmentalisation of cells. I have come across the following statement:
The nucleus and the ...
6
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2answers
224 views
Can we dilute PureExpress Cell Free Mix to increase number of reactions?
Since the PureExpress Cell Free mix is so expensive, I was wondering if it might be possible to just dilute the mix to increase the number of reactions we need to use.
From this image I found:
It ...
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0answers
15 views
Why do molecular generation models maximize “penalized logP” as a measure of drug-likeliness?
I found that Lipinski's rule of five states that Log P (octanol-water partition coefficient, lipophilicity measure) usually should not exceed 5.
Many papers about drug discovery machine learning ...
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0answers
28 views
How to incerase the efficiency of co-immunoprecipitation?
immunoprecipitation recently. My main problem is i am getting very clean and my target protein only (as you see in the lane 3,4,5,6) but when i reduced the NP40 concentration to 0.05 percentage i ...
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0answers
25 views
Low Density Lipoprotein (LDL) in the endosome
I am learning vesicular transport of LDL endocytosis. I understand as pH of the endosome is lower (around 6.0) than cytosol (around 7.2) due to the H+ pump, so LDL disassembles from the LDL receptor ...
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1answer
76 views
How broad are the conclusions that one can make from a heterologous expression experiment?
I am studying whether a neural protein regulates the activity of the protein GSK3. I am using heterologous expression systems where I am overexpressing the neural protein in CHO cells (via ...
3
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1answer
109 views
How can I change the payload of a bacteriophage used to transform E. coli?
I was looking at bacteriophages and how they're used to transform E.coli. While the whole process of how a bacteriophage works makes sense theoretically, I wanted to know how one goes about changing ...
2
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1answer
156 views
Can restriction enzymes (Type II) displace single-stranded binding proteins (SSB)?
Many type II restriction enzymes have been shown to be able to cut ssDNA.
If single-stranded binding proteins are bound to single-stranded DNA (ssDNA), does this prevent restriction enzymes which cut ...
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1answer
77 views
Does anyone have a tutorial or some guide to get started with Rosetta protein modeling and design tool? (I am a Mac user) [closed]
I have been trying to get started with Rosetta protein modeling and design software bundle. I searched online but the tutorials in the Rosetta website are difficult follow. I have the binaries already ...
8
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1answer
225 views
Why is electroporation not the prefered choice transforming for Mammalian Cells (HEK)?
I see a lot of folks using different techniques for transforming mammalian cell (specifically HEK) instead of doing electroporation like I see with E.Coli (bacteria). Is there a reason for this ?
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19 views
Does the two-state gene expression model apply to constitutive genes?
Wikipedia defines constitutive genes as
a gene that is transcribed continually as opposed to a facultative gene, which is only transcribed when needed.
I don't have a strong theoretical background ...
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1answer
37 views
Is the phrase “transmembrane segment” equivalent to the transmembrane domain of a protein?
I am reading the Handbook of Neurochemistry and Molecular Neurobiology and I am learning about the cell adhesion molecule NCAM2 and I have come across the following:
The overall structure of NCAM2 ...
1
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1answer
15 views
What is meant by “opposing plasma membrane” with respect to cell adhesion molecules?
I am reading the Handbook of Neurochemistry and Molecular Neurobiology and I am learning about cell adhesion molecules (CAMs) and I have come across the following:
CAMs are involved in homo‐ or ...
11
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2answers
263 views
Can I use multiple bicistronic RBS sequences in a synthetic biological circuit?
The bicistroninc RBS sequences (BCDs) developed by Mutalik et al. [1] aim to remove context sensitivity from translation and therefore ensure more predictable gene expression. However, I have been ...
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0answers
19 views
Does Invertase catalyse the hydrolysis of other sugars other than sucrose?
I recently did a lab where we tested out the substrate specificity of Invertase on different types of sugars such as sugar alcohols and disaccharides, measured by the absorbance of red using a ...
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0answers
21 views
Surface Plasmon Resonance (SPR) Predicting Molecular Weight Limit?
Just asking, can we predict how low molecular weight (LMW) analyte our SPR system can detect based on our LOD (limit of detection) and our sensitivity on another protein detection? So right now our ...
2
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1answer
23 views
Confusion regarding the Kir2.1 inward-rectifying potassium channel
I was trying to find out more about gustation and the transduction of sourness when I came across the supposed inward-rectifying potassium channel $\ce{K+_{ir} 2.1}$. Here's the thing, despite being ...
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0answers
9 views
Is protein intake required for the formation of new myosatellite cells?
Hypertrophy of muscle fibers requires an adequate (significant, really) intake of protein, but what about just forming new myosatellite cells as a response to a stress applied to the muscle? Does that ...
3
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1answer
57 views
Interaction of charged triphosphates with the ATP binding region in proteins
DNA-histone interactions involve positively charged amino acid side chains groups neutralizing the negatively charged phosphate of the sugar-phosphate backbone. Is the same true for ATP binding sites ...
2
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2answers
87 views
How to assemble three 60mer nts by pcr?
Good morning,
I am new to molecular biology. The question might be silly but i would like to know the answer. I have three 60mer single strand synthetic oligonucleotide. Namely Tag 1 - 3. My goal is ...
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0answers
43 views
Which enzymes use ATP?
It is well known that there are many enzymes which use ATP in their function, and any enzyme that work against an energy gradient need to have that energy supplied from somewhere, but just as well ...
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1answer
36 views
Electrochemical and concentration gradient confusion [closed]
I am learning membrane transport and learnt that there are 2 gradients that determine passive and active transport.
If the concentration of a cation is higher in the cytoplasm side (hence ions should ...
0
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1answer
70 views
DNA complementarity against reverse complementarity [duplicate]
I am sorry to bother with this question (i study genetics for about few hours, because I need to understand my data) and I am really confused about these two terms, because I dont know if the books ...
4
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1answer
103 views
Introns and miRNA
From this paper, it is stated that some introns may contain genes coding for miRNA, miRNA is essential in regulating gene expression by pairing with RNA, hence disrupting regular translation.
From ...
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0answers
33 views
Dealing with heterochromatin during DNA replication
Heterochromatin are present along the chromosome (uncoiled state). With the highly-condensed structure relative to euchromatin, RNA polymerase cannot get into the DNA base pairs in heterochromatin and ...
0
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0answers
35 views
Flip-flopping in plasma membrane
Flip-flopping of lipids (and in proteins it is impossible) in plasma membrane is rare due to high energy barrier (video ref). However, it is an important mechanism since it allows asymmetric ...
7
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1answer
141 views
Why can't certain nucleoside analogs be used to label bacterial RNA?
A 2020 publication in Nucleic Acids Research 1 includes the following passage:
A variety of nucleoside analogs have been developed for metabolic RNA labeling in various eukaryotic cells (9–16). Among ...
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Western blotting: what common tweaks do you make to the protocol and why would you make membrane cuts?
I'm building an app which helps researchers plan out Western blots, record protocol tweaks and receive warnings in the app if:
you're trying to multiplex antibodies of similar molecular weight on the ...
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1answer
37 views
Can caffeine act as viagra
Recently I studied that cAMP which is a 2° messenger can cause various physiolocal changes under the action of proper hormones. Now cAMP is degraded by phosphodiesterase and this ceases the actions. ...
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0answers
20 views
Question on cellulose synthase
How does the rosette structure of synthase cause the cellulose synthesized to be aligned next to each other?
I just cannot visualise how a rosette structure can be able to cause the celluose to be ...
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36 views
Question about : RNA interference - small-interference RNA AND about Rab-protein in vesicles
First Question :
RNA interference - small-interference RNA
in the textbook -Essential cell biology By Bruce Alberts, Dennis Bray, Karen Hopkin
this Figure is Shown
my question is : if the foreign RNA ...
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27 views
Need help designing an enzyme immunoassay (EIA)
I have a FITC-labelled DNA sequence and an anti-FITC antibody, however this antibody is not conjugated with HRP or any other enzyme. I work in a lab with limited finances, so is there a way I could ...
0
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1answer
26 views
Which landmark paper first described the differentiation of T-cells?
T-cells are distinguished from B cells in part by their locus of differentiation/maturation (thymus). This is textbook knowledge, but I was wondering which particular person or people were responsible ...
