Prewitt

i agree with the above. best would be to have a strain without the protein (then you can see what it does anyway, there are better means for that) and then put a plasmid with the chormome in it. you have to show that these are equal if you want to make conclusions about in vivo. then you can make a strain with a mutated plasmid and can compare these. i'd suggest in vitro studies before that though. translate it in vitro translations n stuff. better to start basic that complex.

also that is not entierly true. it is pretty often just ONE cell that makes a cancer, since most are monoclonal. the point is that cancerous cells arise all the time and degrate by various means, being limited by telomorases being probably one mean, and nothing happens. but if just ONE cell does survive that is cancerous, you will get a solid cancer. that is the way it happens. it's all just statistics. there is a large number of cells that mutate. few of these mutations are sufficient for cancer. and few of these survive to actually cause cancer. but if ONE does, you get cancer. normally that numbers are so small that you probably don't get cancer. once on of the numbers increase, the probability does too. it just matters if one cell in the end survives.

that is...well...not completely correct. there is a reason that cells do not have a telomerase. if a normally non-proliferative tissue get by whatever means proliferative in a cancerous way, it's proliferative potency is limited to the normally lacking telomerase. if you artifically give all your cells a telomerase, cancer prevalence among your cells will drastically increase. may i ask what relation you have to the topic? are you a student?

last thing i've heard were experiments in the states who thought they could and some patients died. because the vector worked well in just a part of the patients. also the problem is that it's just modified viruses afaik and you can't make them specific for any cell you like.

sadly enough, this is not necessarily true. there are some problems in science that can not be solved by solely working on them. eg. the development of a fusion reactor. there are profound problems that might be solved by the discovery of a totally new technique. unfortunatly those discoveries happen by accident rather than systematic research. therefore in vivo cell targeting might be invented one day, but might also not at all. electroporation is a rather old in vitro technique for dna uptake that does not contribute the the mentioned problem of specific targeting of in vivo cells which is without doubt a necessity. therefore to my mind all thought about the topic are pure science fiction. and it'll stay that way until something like a revolutionary discovery is made. as i already said this discovery can occur but doesn't have to. you can but a million highly skilled scientists in a garden eden of research - they might work several hundred years and still not make the discovery - that's the way my math teacher used to explain this kind of discovery.

the problem is not really to manipulate the gen but to target cells specificly. there have been deaths because artificial viruses targeted the wrong cells in some patients. imo that is the major problem. once you can specificly target your cells of intereset in vivo (meaning them and only them) medicine will enter a new century.

telomere length is just a thesis. there is probably a lot more to limitations of the lifespan of cells than just telomeres. besides - you cannot manipulate genes that easily in living humans. if you could - hellooooooooo nobel prize.

i'd suggest reading this if what you find with google ain't enough: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=AbstractPlus&list_uids=10658174&query_hl=1&itool=pubmed_DocSum

it is appearently the correct solution values are ΔH0 from Breslauer 1986 AA 9,1 AC 6,5 AG 7,8 AT 8,6 CA 5,8 CC 11,0 CG 11,9 CT 7,8 GA 5,6 GC 11,1 GG 11,0 GT 6,5 TA 6,0 TC 5,6 TG 5,8 TT 9,1 5’-AA-3’: 9,1 3’-TT-5’ --> 5’-TT-3’: 9,1 5’-AT-3’: 8,6 3’-TA-5’ --> 5’-AT-3’ 5’-TA-3’: 6,0 3’-AT-5’ --> 5’-TA-3’ 5’-CA-3’: 5,8 3’-GT-5’ --> 5’-TG-3’: 5,85 5’-GT-3’: 6,5 3’-CA-5’ --> 5’-AC-3’: 6,5 5’-CT-3’: 7,8 3’-GA-5’ --> 5’-AG-3’: 7,8 5’-GA-3’: 5,6 3’-CT-5’ --> 5’-TC-3’: 5,6 5’-CG-3’: 11,9 3’-GC-5’ --> 5’-CG-3’ 5’-GC-3’: 11,1 3’-CG-5’ --> 5’-GC-3’ 5’-GG-3’: 11,0 3’-CC-5’ --> 5’-CC-3’: 11,0

hm. might be, i'll check it out...thanks for the input

i said there aren't distinct borders. I've been working in the institute of biochemistry here for 7 years and in a molcular biology lab for 4 years now. the difference lies on the point of focus, not on the methods itself.

*bump* anyone got a clue or a theory?

you can listen to me or not but if i post something you can be sure i am quite certain of it. to underline my point you can consider the most famous molecular biology book with is the alberts ("cell") it exculsivly deals with dna, rna and the cell's work on these. still can't name an english textbook for pure biochemistry though. don't let the school books fool you: they're named "biochemistry" but include both. why? cuz they were started back in the days of good old protein chemistry, before taq and the like made their way into the lab.

i didn't read all the posts in this thread so maybe i'm missing something. but there is a difference. biochemistry focuses on proteins. molecular biology on nucleotides. you could also say biochemists use acrylamide gels and molecular biologists use agarose gels most of the time - metaphorically spoken. there is no distinct border of course. to prove this thesis, i cannot name you any english textbooks because as far as it involved english literature i just read protocol books (eg. the red book) but if you consider a more schoolish kind of literature the difference is apparent (eg. books of the "experimentator" line - it's german i know but see above )

just did that twice...but 1.how did you notice you protein was degraded? --> see if buffer ph and omsomlarity are optimal 2.to avoid contamination: wash more. 3.gst is a dirty thing. it simply is.

how bout an ezymatic assay? it's abit old fashioned but should give figures if that is what you are looking for.

i don't have much experience with antibody production but it seems quite reasonable titres increase. you have plenty of epitopes on the carrier protein. but since only the ones for your hapten-like (appearently) protein really matter i'm not sure it gives you better antibodies. but that's just speculation.

i'm currently writing a piece of c++ code for improved primer selection for our lab. nothing fancy really. lately i was annoyed by the incorrect melting temperature calculated by 4+2 rule therefore i upgraded to nearest neighbour method. i read the papers by breslauer, santalucia, sugimoto and the rather crappy one by panjkovich - but i'm not really into the subject matter. question is: why are there only ten watson crick interactions that all authors provide thermodynamic data for? They provide data for AA, AT, TA, CA, CT, GA, GT, CG, GC and GG (with complementary nucleotides) but not for AC, AG, CC, TC, TG, TT. I wouldn't really care, but since i want to calculate it right: do i just skip those pairs in calculating the integrals?