6 replies to this topic

[boka]

Hi everyone,

I used to drop by these forums once in a while and have a bit of a browse, but have never actually posted until now.

I am a third-year science student and I need some help making sense of some parts of a journal article (Here is a .PDF file of the full article). I understand the general concept and findings of the study but I have never taken any genetics courses at uni and hence I am somewhat weak in my understanding of some of the terminology used in the materials and methods section. I currently don't have access to the biological sciences library as it is closed for the easter public holidays so I am without any reference texts.

I am to give a fifteen minute presentation about a week from now and would really appreciate it if someone with a better mastery of genetics can briefly describe/explain the techniques that were used and how they relate to the figures presented in the article.

Thank-you!  

[methylnick]

hi,

which terminology were you unfamiliar with within the materials and methods section?

Nick

[boka]

Thanks for replying, methylnick

There's quite a few acronyms/abbreviations and also techniques themselves for which I don't have any knowledge of eg. from the lentiviral expression section, what is TOPO-cloning? What is pLenti6 V5-D and pLP1, pLP2, and pLP/VSVG? What's the difference between 293FT cells and HEK293 cells? Why would you want to determine the number of Blasticin-resistant colonies?

My goal is to make some sense of what's being shown in figures 2 and 3.

Edited by boka, 26 March 2005 - 10:09 PM.

[methylnick]

boka, on Mar 26 2005, 11:07 PM, said:

Thanks for replying, methylnick

There's quite a few acronyms/abbreviations and also techniques themselves for which I don't have any knowledge of eg. from the lentiviral expression section, what is TOPO-cloning? What is pLenti6 V5-D and pLP1, pLP2, and pLP/VSVG? What's the difference between 293FT cells and HEK293 cells? Why would you want to determine the number of Blasticin-resistant colonies?

My goal is to make some sense of what's being shown in figures 2 and 3.

<{POST_SNAPBACK}>

TOPO cloning is a method of getting the DNA of interest, in this case mutated forms of the gene, into a plasmid for delivery and expression.

Anything denoted with a p is a plasmid and it's name.

Not too sure about the different 293 cells.

I think the whole gist of the figures is to show the mutations and that the E1093K mutation has some functional effect!

Hope it helps!

Nick

[boka]

Thanks Nick, it's amazing how a short explanation can go a long way

Just one more question to be absolutely clear, is E1053K just a label they put on the mutation, or does it have some sort of significance in the way of describing where the mutation occurs or what is changed?

[Simonsays]

E1053K means that the glutamic acid (E) in position 1053 is changed for a lysine (K).

Simon

[boka]

That makes sense, thanks Simon