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Treatment of RNase with 8M urea but without adding mercaptoethanol would?
Wiki User
∙ 13y ago
Urea itself will only break non-covalent bonds, breaking any protein into its basic subunits, polypeptide chains of amino acids. Without mercaptoethanol this would be the end result, and from this you could deduce whether the protein was a dimer, tetramer, etc. Mercaptoethanol is an oxidizing agent which is used to oxidize the disulfide bridge formed between cysteine residues. Oxidation of these sulfur atoms results in the disulfide bridge being broken, further reducing the protein (now polypeptide chains) into its smaller subunits. If successful, all of the collected protein remains should equal the total molecular weight observed in the protein in its entirety.
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∙ 13y ago
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What does RNase solution do in the Isolation of genomic DNA from Negative bacteria?
Digests RNA molecules
What did Sidney altman contribute to science?
In the early 1980s, Sidney Altman discovered that ribonucleic acid (RNA) molecules can act as enzymes. This disclosure, independently and concurrently made by Thomas R. Cech of the University of Colorado, broadened our understanding of the origins of life. Before this discovery, it was believed that all enzymes were made of protein and that primitive cells, therefore, used proteins to catalyze biochemical processes. Now, it appears that RNA may have acted as a catalyst . Altman and Cech's work has not only had a "conceptual influence on basic natural sciences," according to the Royal Swedish Academy of Sciences, but in addition, "the discovery of catalytic RNA will probably provide a new tool for gene technology, with potential to create a new defense against viral infections." As a result of their findings, Altman and Cech were jointly awarded the 1989 Nobel Prize for chemistry. Altman was born in Montreal, Quebec, on May 8, 1939, the second son of Victor Altman, an immigrant grocer, and Ray Arlin, who before her marriage worked in a textile mill. He attended West Hill High School in Montreal and the Massachusetts Institute of Technology, from which he graduated with a bachelor of science in physics in 1960. Between 1960-1962, he was a teaching assistant in the Department of Physics at Columbia University, while he waited for a suitable position in a lab. Around this time, Altman switched from physics to the newly emerging interdisciplinary field of molecular biology. He moved to the University of Colorado in Boulder in late 1962 to work as a research assistant under Leonard S. :mb Lerman. :mx He was mainly preoccupied with studying the replication of the T4 bacteriophage , a substance that infects bacterial cells in much the same way as a virus infects human cells. Altman received his Ph.D. in biophysics in 1967. After graduation, Altman briefly worked as a research assistant in molecular biology at Vanderbilt University before winning a grant from the Damon Runyon Memorial Foundation for Cancer Research. This permitted him to work as a research fellow in molecular biology at Harvard University. From 1967 to 1969, working under the biochemist Matthew Meselson, he continued his research into the genetic structure of the T4 bacteriophage. His receipt of the Anna Fuller Foundation Fellowship in 1969 enabled him to transfer to the Medical Research Council Laboratory of Molecular Biology in Cambridge, England, to work with molecular biologists Sydney Brenner and Francis Crick. The latter, in partnership with James D. Watson, discovered DNA's double-helix structure in 1954. It was clear to scientists that genetic information is carried by DNA (deoxyribonucleic acid ) into a cell's nucleus. In the cytoplasm (the substance inside the cell wall, surrounding the nucleus), the genetic code is copied into RNA. It is then converted into proteins , which are built of chains of amino acids. Altman originally intended to study the three-dimensional structure of transfer RNA (tRNA), which is a small component of RNA that transfers amino acids onto a growing polypeptide chain as proteins are made. Much of the breakthrough work in this area had already been accomplished, however, so Altman decided to switch his attention to the transcription of tRNA from DNA. He found that the DNA from which tRNA is produced is not directly copied into tRNA but first undergoes an intermediary stage when it becomes a long strand of what is called "precursor RNA ." This is composed of a strand of tRNA with additional genetic sequences at each end which are somehow later removed before it becomes tRNA. While still working at the Medical Research Council in Cambridge, Altman studied the tRNA genes of the Escherichia coli (E. coli) bacterium, to which he added toxic chemicals. Subsequent mutations in the tRNA enabled him to isolate precursor tRNA from bacterial cells. Altman discovered that the additional sequences at each end of the strand of precursor tRNA were removed by something in the cells of the bacteria, probably an enzyme. The scientists found that the enzyme, named ribonuclease P (RNase P) , would only cut off the extra sequences at a precise point. When he returned the United States in 1971, Altman joined Yale University's biology department as an assistant professor. In 1972, he married Ann Korner. They have a son, Daniel, and a daughter, Leah. Altman was promoted to associate professor in 1975. In 1978, he published the results of an experiment carried out by one of his graduate students, Benjamin Stark. It demonstrated that RNase P was at least partially composed of RNA, which meant that RNA itself played an integral part in the activity of the enzyme. This finding was highly unorthodox, as it was then presumed that enzymes are made of protein, not nucleic acids. In 1980, Altman attained a full professorship of biology at Yale. The following year, Cech at the University of Colorado published independent results similar to Altman's. Cech discovered that the precursor RNA from the protozoan Tetrahymena were reduced to their final size as tRNA without the assistance of protein, and suggested that the precursor RNA catalyzed this itself. His findings lent weight to Altman's. Cech's use of the word "catalyst" to describe the action of the RNA was questioned, however, because rather than just speeding up a reaction, it used itself up in the process. Three years later, by which time Altman had become chairman of Yale's biology department, his colleague, Cecilia Guerrier-Takada, was testing the catalytic activity of RNase P. She discovered catalysis even in the control experiments that used the RNA subunit of RNase P (the M1 RNA) but which contained no protein. Altman was able to prove that the M1 RNA demonstrated all the classical properties of a catalyst , especially as, unlike that studied by Cech, it remained unchanged by the reaction. This removed the last shadow of a doubt that RNA could act as an enzyme. In 1984, Altman became a naturalized American, but retained his Canadian citizenship. From 1985 until 1989, as the dean of Yale College, Altman established a greater role for scientific education in all of Yale's curriculums. In 1989 he and Cech jointly received the Nobel Prize for chemistry for their discovery of RNA's catalytic ability. Their work put an end to the conundrum regarding proteins and nucleic acids which had long mystified scientists. They had been unable to discover which came first in the development of life, proteins or nucleic acids . Proteins catalyze biological reactions, whereas nucleic acids, such as RNA, transport the genetic codes that create the proteins. Altman and Cech proved that nucleic acids were the building blocks of life, acting as both codes and enzymes. High hopes exist for the practical applications of their discovery, which was described by the Nobel Academy as one of "the two most important and outstanding discoveries in the biological sciences in the past 40 years," the other being Crick and Watson's discovery of DNA's double helix structure. If RNA enzymes are able to cut additional sequences of tRNA from a strand of precursor tRNA , doctors could possibly use RNA enzymes to cut infectious RNA from the genetic system of a person with an infectious viral disease. Research into this field is ongoing and, if fruitful, could contain the key to curing viral infections such as cancer and AIDS. In addition to receiving the Nobel Prize, Altman was honored with the Rosentiel Award for Basic Biomedical Research in 1989. He is a member of the National Academy of Sciences and the American Society of Biological Chemists, of the Genetics Society of America, and is a fellow of the American Association for the Advancement of Science. He holds honorary degrees from the Université de Montréal, York University in Toronto, Connecticut University, McGill University, the University of Colorado, and the University of British Columbia. In 1991, he was selected to present the DeVane lecture series at Yale on the topic "Understanding Life in the Laboratory." Altman has held a number of other part-time positions in addition to his full-time academic positions, including associate editor of Cell from 1983 to 1987, member of the Board of Directors of the Damon Runyon-Walter Winchell Fund for Cancer Research, member of the Board of Governors of the Weizmann Institute of Science, member of the Scientific Advisory Board of Bio-Méga, Inc., and Special Consultant to the Pathogenesis Corporation in Seattle.
What is the purpose of RNA extraction?
RNAse destroys the RNA and hence RNAse contamination is a problem in RNA extraction as it breaks down RNA. RNAse enzyme is removed by using RNAse inhibitor or precautions like wearing of gloves, autoclaving tips , using RNAse free water/DEPC treated water is done while performing RTPCR
Role of RNase in DNA purification?
RNAase remove RNA from the solution as we need DNA only. RNAse cuts down the RNA strand.
What is the role of DEPC in RNA isolation?
It inactivate the RNase and prevent RNA to denature.
What does RNase solution do in the Isolation of genomic DNA from Negative bacteria?
Digests RNA molecules
What is superasin?
An RNAse inhibitor, made by Ambion http://www.ambion.com/techlib/resources/superasein/
What is the role of TRIreagent in RNA isolation?
Break open the cells, stabilize RNA, inhibit RNAse.
Which is the enzyme without protein?
Ribozyme is an enzyme made of ribonucleic acid (RNA).
What epidermal cell defends skin against microbes?
i don't know about chemicals but epidermis cells produce RNase enzymes to destroy microbial genomes
What has the author Katrin Trautwein-Fritz written?
Katrin Trautwein-Fritz has written: 'Construction of an improved expression system for bovine pancreatic ribonuclease A and construction and characterization of RNase A mutants' -- subject(s): Gene expression, Proteins, Recombinant proteins, Ribonucleases, Synthesis
What enzymes are in the mouth?
The saliva in mouth has following enzymes:salivary amylase or ptyalin (breaks down carbahydrates)lingual lipase (enzyme for fats digestion)lysozyme (enzyme that kills bacterial cell)salivary ribonuclease (RNase)Deoxyribonuclease (DNase)
Why RNA extraction is difficult?
RNA itself difficult to handle because of its unstable nature. Unlike DNA, RNA has free 2'-OH group in their ribose sugar that make them highly reactive. Other than this, RNAse contamination is everywhere (during isolation RNAase from our skin can kill RNA).
How does a plant self pollinate?
By having the pistils form a few days after the stamens do.In some plants, a toxin called S-RNase poisons the pollen tube if the pollen and pistil are too closely related, thus preventing inbreeding. Credit: Nicolle Rager Fuller, National Science Foundation
