what plasmid is capable of integrating into bacterial chromosome?
371069
Well, at the least conjugative plasmids are known to occasionally integrate into bacterial chromosomes. I think some others are, but if I had to pick just one, that would be it.
584734
I gave a lecture on this recently and have had first hand experience with this technology in a lab, so let me give you a brief and easy summary to take with you about this topic about Recombinant DNA Technology:\n\nIn order to clone you need 2 things: Bacterial DNA (the circular form called a "plasmid"), and the DNA you want to clone (be it human, mouse, plant, other species etc)\n\nThen you will need to cut the Bacterial DNA with enzymes called "endonucleases" that work like scissors. (So imagine a circle and I take a pair of scissors and cut the circle at some point in it, b/c I want to insert the foreign DNA (non native DNA) where I had cut.) I will have to do the same with the foreign DNA and have endonucleases cut OUT the particular piece i want cloned. \n\nThen we can put the two pieces together, the cut circular plasmid DNA from the bacteria, and the CUT out piece of foreign DNA. \n\nWhen we put the two pieces together it is called "ligation", and it is done as a reaction in a tube with enzymes inside that "glue" the "sticky ends" of the bacteria DNA to the foreign DNA. \n\nAfter the reaction normally done in a 16 deg. Celsius water bath, we now will have "circular" plasmids with our Foreign DNA inserted into the bacterial DNA (the circle DNA)\n\nThen we would re-insert this DNA back into the bacterial host (that the circle DNA first came out of) in a process called "transformation", this is where the broth from the water bath we just discussed is grown in a growth medium at 37 deg. celsius so that the bacteria can grow and produce MORE of those circle plasmids with our foreign DNA inserted into it. \n\nThen we would "plate" these onto agar plates, and let them grow in incubators at 37 deg. Celsius over night until we get small colonies of bacteria on the plate. \n\nEach "mound" of bacteria (small like white dots on the plate) contains millions of copies of our gene (inserted in the circular plasmid DNA of the bacteria). \n\nNow we can take these small colonies, and harvest them further, by putting them in large Erlenmeyer flasks to be grown in the same "growth broth" over night on a shaker for 37 deg. celsius. \n\nAfter of which we can harvest, and purify that culture so that only the circular plasmids remain (this is a long and convuluted process so we won't go into it in detail here), and from there you can store those in a 4 deg. Celsius fridge and keep it in stock for further research purposes. \n\nHope that helps!
852508
NaCl needs to be at the correct concentration, as it is needed to ensure that enzymes etc. work correctly.  It allows the cells to have the correct concentrations of sugars etc by influencing water entry by osmosis.\n\nA cell can take up a linear plasmid, but probably won't be able to propagate more copies if is isn't annealed, unless the correct DNA Ligase enzymes happen to be available in the cell.
452878
From my understanding tDNA is important, at very least for tagging but I believe it is also important for the transfer of the intesert into the A. thaliana genome.\n\nYou can still detect the insertion of the transgene because you would usually insert an antibiotic resistance gene with the gene of interest. You can then use this to select for plants that have the gene inserted.\n\nThen, to find out where in the genome the transgene has inserted, you need to map it. There are many techniques for doing this, but if you know the sequence of your gene of interest or the antibiotic resistance gene, you should be able to use fragments of the genes as probes on gene libraries etc etc etc, there are probably also fancy ways you could use genomic to achieve this too.
212530
You need a DNA plasmid with a few things.  First a origin of replication, so the plasmid can be replicated.  Second, an antibiotic resistance marker to issolate only bacteria containing the plasmid.  Third you need a bacterial promoter with a consensus kozak sequence followed by your gene of interest in frame.  Usually this gene is followed by or preceded by glutathione-S-transferase (GST) to facilitate purification of the protein).  Also usually the promoter contains a repressor region LacI, which means the gene is turned off normally, unless you add lactose or IPTG (a synthetic lactose), which turns on gene transcription and translation.  This is routinely done in labs nowadays.
244250
You have to identify two sequences that are >6kb apart from one another that flank the sequence you are attempting to amplify. Better yet, you can get you a well understood cloning vector (plasmid) that already has primers develped for it that flank the polyrestriction site region (ie. puc19 [see link below], find the needed restriction enzyme sites that flank your desired 6kb sequence and clone the DNA into the vector. Be sure to check your sequence for any internal restriction sites. This may require you to do a partial digestion and recovery.
744973
Prokaryotic  proteins -\n1. Cro and repressor proteins\n2. LacI purine repressor proteins (PurR)\n3. FokI restriction endonuclease (DNA-recognition region)  \n4. Gamma-delta recombinase protein \n5. Hin recombinase protein\n6. Trp repressor protein \n7. Diptheria tox repressor \n8. Catabolite gene activator proteins (CAP)   \n\nEukaryotic proteins -\nHomeodomain proteins \n2. RAP1 protein \n3. Prd paired protein  \n4. Tc3 transposase protein \n5. TFIIB family  \n6. Interferon regulatory factor \n7. Transcription factor family \n8. ETS domain family \n9.TATA Box-Binding Proteins \n\nThese are some protiens associated with DNA
718212
dr d is correct.\n\nIt is transfer of genes within a generation, as compared to transfer from the parental generation to the offspring generation; that is vertical gene transfer between generations.\n\nIf genes are picked up from a cell in some way, and put into other cells (not involving cell division), then the genes have been transferred "horizontally".\n\nAn example is when a bacterium carrying a certain plasmid grows a "sex pilus", which then connects with the cell wall of a bacterium that does not carry that plasmid.  It will inject a copy of the plasmid, via the sex pilus, and the recipient has then become a carrier, too.  These plasmids often carry extra genes, as well as those coding for this ability.  These phenomena are sometimes considered to be similar to sex, but are really more like catching a virus!
866554
There are more than two, depending on specifically what you are doing.\n\nGenerally transformation is of bacteria. The two major methods for doing this are plasmid transformation and phage transformation. (see link below)\n\nThere are additional methods used on eukaryotic cells.\n The Shotgun approach coats minuscule particles of gold with the DNA you want transferred and shoots them into the cells. Insertion is random.\n\n Some plants are amenable to agrobacterium mediated transformation. This is much more precise than the shotgun approach. (see link below)
843667
How is Recombinant DNA made? \nThere are three different methods by which Recombinant DNA is made. They are \nTransformation, Phage Introduction, and Non-Bacterial Transformation. Each \nare described separately below. \n\nTransformation\nThe first step in transformation is to select a piece of DNA to be inserted \ninto a vector. The second step is to cut that piece of DNA with a restriction \nenzyme and then ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable \nmarker which allows for identification of recombinant molecules. An antibiotic \nmarker is often used so a host cell without a vector dies when exposed to a certain \nantibiotic, and the host with the vector will live because it is resistant. \n\nThe vector is inserted into a host cell, in a process called transformation. One \nexample of a possible host cell is E. Coli. The host cells must be specially \nprepared to take up the foreign DNA. \n\nSelectable markers can be for antibiotic resistance, color changes, or any other \ncharacteristic which can distinguish transformed hosts from untransformed hosts. \nDifferent vectors have different properties to make them suitable to different \napplications. Some properties can include symmetrical cloning sites, size, and \nhigh copy number. \n\nNon-Bacterial Transformation\nThis is a process very similar to Transformation, which was described above. The \nonly difference between the two is non-bacterial does not use bacteria such as E. Coli \nfor the host. \n\nIn microinjection, the DNA is injected directly into the nucleus of the cell being \ntransformed. In biolistics, the host cells are bombarded with high velocity \nmicroprojectiles, such as particles of gold or tungsten that have been coated \nwith DNA. \n\nPhage Introduction\nPhage introduction is the process of transfection, which is equivalent to transformation, \nexcept a phage is used instead of bacteria. In vitro packagings of a vector is used. \nThis uses lambda or MI3 phages to produce phage plaques which contain recombinants. \nThe recombinants that are created can be identified by differences in the \nrecombinants and non-recombinants using various selection methods.
598653
To try and answer the question, rather than tell you all about bacterial cell walls again... I think that yes, the wall is an integral part of the bacterial cell membrane. If it was a generic question on the "functions of carbohydrates in cell membranes", then I'd expect an overview of how different carbohydrates are used in different ways, in different cell membranes, including the bacterial ones.\nIt's a bit unfair of your teacher to fail everyone, however - if no one mentioned it, it sounds as if the question wasn't asked clearly. \nIs anyone in the class thinking of challenging him/her on it? Good luck!
154824
This seems like a hard question but if I understand it right it is asking you why glucose is used in isolation of plasmid DNA.  From my deduction it is simply a growth medium for the bacteria providing enough bacteria with the plasmid.  Then the lysozyme is added to break down the cell walls of the bacteria to begin the isolation of plasmid DNA.  This source may help: http://www.bio.com/protocolstools/protocol.jhtml?id=p571
392177
some bacteriophages were radiolabelled with radioactive phosphorous and sulhur. the sulphur bound to the external  protein coat hile the phosphorous bound to the nucleic acid inside. now, these pahges were allowed to infect a bacterial colony. the infected bacterial colony was then separated and subject to centifugation. it revelaed that while the radioactive sulphur was leftbehind in the phages, the bacterium had contracted the radioactive phosphorous. it was suggested that this was because once the phage infected the bacterium, they transfered their DNA into the bacterium. this is called transformation. this has now been verified by other methods. transformation is a very important method for genetic mutation studies.
244592
Certain ligases which cut DNA at known sequence points are used to excise the gene of interest from a particular strand of DNA. This excised (or cut-out) gene is then incorporated into the DNA of a bacterium.  The bacterium, in the process of replication, will add the gene into its own plasmid and produce the protein. Alternatively, I believe viral vectors are also used to introduce genes into a particular host "factory" cell that then produces the required protein. Not totally informed on that part, though.
638715
Stain with ethidium bromide (careful, its a carcinogen) and photograph under uv light.\n\nMeasure the migration of the bands and those of the size reference.  Plot the migration of the size reference on semi-log paper and use the resulting curve to detemine the size of your plasmid fragments.
935437
Yes, through genetic splicing as long as an intron is inserted as well.
702124
You can find practical information from Qiagen website if you use their kits which are quite popular these days.  \n\nIt depends on which methods you use for isolation.  The alkali lysis (see link below) was commonly used to rupture bacteria.  Plasmid DNA (smaller in size and more soluble) is separated from baterial genomic DNA (larger and less soluble).  The plasmid DNA is then separate from proteins by phenol extraction where the proteins are removed to the phenol phase while plasmid DNA stays in the aquous phase.  The plasmid DNA is then precipated with ethanol to complete the purification.\n\nQiagen's kit and others overcome the use of phenol (which is toxic and smelly) by using a solid phase that bind DNA to seperate the contaminating proteins.  The DNA is later eluted from the solid phase to complete the purification.\n\nHere are additional links:\nhttp://biotech.biology.arizona.edu/labs/DNA_isolation_plasmid.html\nThe concept of alkali lysis method of plasmid DNA isolation is shown here:\nhttp://a32.lehman.cuny.edu/molbio_course/plasmid_protocol.htm