Two different strains of TMV have different forms of a viral protein that can be distinguished by whether they cause large or small spots on the tobacco leaves. [12] mRNA is selectively reverse transcribed using oligo-dT primers that are the reverse complement of the poly-adenylated tail on the 3' end of all mRNA. RNA polymerase proceeds at about 50 nucleotides per sec. Since each deoxyribose in a DNA chain is coupled to one of the four nitrogenous bases (G, A, T, or C), these long chains can carry information. However, it differs from DNA and RNA polymerases in points 1 and 4. However, priming is random and hairpin hydrolysis leads to loss of information. 204A/170A sec-1 = 1.2 sec per initiation, or 0.83 initiations per sec. See this is the correct answer. The normal version of the gene that can be mutated to cause the disease is known. It is possible to reconstitute TMV in the test tube by mixing pure viral proteins and pure viral RNA. Okay, so the correct answer is option number A. I hope you understand. Transcription: Promoters and Terminators. The older, template strand is fully methylated and the post-replication repair pathway exploits this to repair the newly synthesized strand. For the best experience on our site, be sure to turn on Javascript in your browser. Often, when the word DNA is used without further clarification, it refers to gDNA. [10] The AMV reverse transcriptase from the avian myeloblastosis virus may also be used for RNA templates with strong secondary structures (i.e. 5'Kpn 500 Bgl Bgl 1500 Kpn, ---------------------> *--------------------------------------->, <--------------------* <---------------------------------------, , 5' Bgl Kpn, *--------------------------------------->, | | | | | | | | | | | | | | | | | | | | | | | | |. The substrates (in the direction of synthesis) are ribonucleoside diphosphates, which are added with the release of phosphate as a product. DNA can copy itself in a process known as replication, using DNA polymerase. E.g. Everything you need for your studies in one place. In this question it has been asked Which of the following statement is true bob Seger DNA library C C D N. A. b. The 5' GUN codon can be matched with anticodons 3' CAI + 3' CAC, or 3' CAG + 3' CAU. DNA is made up of four different building blocks, called nucleotides, which are each made up of one of four nitrogenous bases demonstrated in Figure 1. Experts are tested by Chegg as specialists in their subject area. Much less likely are two-base changes from GAA to GTG, GTT, or GTC; and from GAG to GTA, GTT, or GTC. , and this was a key technique in deciphering the genetic code. AUG codons in the interior of the mRNA are eventually positioned at the ribosomal P site and can bind and incorporate only Met-tRNAMet. 94% of StudySmarter users get better grades. (A) It can be amplified by the polymerase chain reaction. Use the human genome sequence to find micro- or mini-satellite sequences that are closely linked to the structural gene. protein from the initiating ATG to the termination codon. A proofreading activity would sever this link, halting synthesis of the polypeptide and releasing it from the mRNA. If RNA is the genetic material, the progeny virus should produce small spots when used to infect a new plant, and the virus should be true breeding in subsequent generations. This is achieved by designing sequence-specific DNA primers that hybridize to the 5' and 3' ends of a cDNA region coding for a protein. A minimum of two transesterification steps are required. Now we have been given four statements about C D N. A. 3) 16 pt. The word genome is an extension of this concept and means the collection of all genes and other information contained inside the nuclei of an organisms cells. 12.2 One of the key signals for cleavage and 3' polyadenylation is the sequence AAUAAA. When scientists transfer a gene from one cell into another cell in order to express the new genetic material as a protein in the recipient cell, the cDNA will be added to the recipient (rather than the entire gene), because the DNA for an entire gene may include DNA that does not code for the protein or that interrupts the coding sequence of the protein (e.g., introns). Necessary cookies are absolutely essential for the website to function properly. Retroviruses, such as Simian Immunodeficiency Virus (SIV) and Avian Myeloblastosis Virus (AMV), use their cDNA to produce mRNA in the host, leading to the production of viral proteins. Beginning with the last codon (Arg) of the wild-type a-globin gene, the 3' end of the wild-type a-globin mRNA has the sequence: 5'..CGU UAA GCU GGA GCC UCG GUA GCA GUU CCU CCU GAC AGA UGG GCC UCC CAA CGG GCC CUC CUC CCC UCC UUG CAC CGG CCC UUC CUG GUC UUU GAA UAA AGU CUG AGU GGG.3'. The term cDNA is also used, typically in a bioinformatics context, to refer to an mRNA transcript's sequence, expressed as DNA bases (deoxy-GCAT) rather than RNA bases (GCAU). University of North Carolina at Wilmington, Masinde Muliro University of Science and Technology. An optimization of this procedure relies on low RNase H activity of M-MLV to nick mRNA with remaining RNA later removed by adding RNase H after DNA Polymerase translation of the second-strand cDNA. An example of this first step from viral RNA to cDNA can be seen in the HIV cycle of infection. This process is called transcription. 4) Score the pyrA+ transductants for fol phenotypes by plating on minimal glucose + folic acid + or - aminopterin. This is false. Then the enzyme polyadenylate polymerase adds a string of 20 to 250 A's to the free 3' end, generating the 3' poly(A) tail. The protected fragment is from the top strand and is 100 nts (a size that can be measured more accurately than the 1300 nt fragment on the polyacrylamide gels used in this analysis). CAN YOU ALSO EXPLAIN WHY THE OTHER ANSWERS ARE WRONG? b) Based on this information how was the Constant Spring allele likely generated? For example, reconstitute a virus with large spot protein + small spot RNA. Hydrogen is also used to construct ATP and GTP. As a result, cDNA will only contain genes that are actively being used by a specific cell or tissue at a point in time. The Japanese film industry is planning to shoot a series of new Mothra v. Godzilla movies. 2) Infect fol- pyrA+ recipient with lysate. Write out the sequence of the first 25 nucleotides in the flow-gram above, starting from the left (Ignore the very short lines. 13.2 a) It starts at AUG (nucleotides 5-7) and ends at UGA (nucleotides 23-25). BioChains cDNA samples are synthesized using total RNA isolation at the facility with modified techniques to ensure consistency. b) All peptides have the same specific activity. This is the process retroviruses use to incorporate into their hosts cells. pyrA+ will grow on both plates, pyrA- cells will only grow on the supplemented plate. 3) Plate mix on minimal glucose plates + folic acid to select for pyrA+. Generally, introns are spliced out, or edited out of RNA in the transcription process before proteins are created. In genetics, complementary DNA (cDNA) is DNA synthesized from a single-stranded RNA (e.g., messenger RNA (mRNA) or microRNA (miRNA)) template in a reaction catalyzed by the enzyme reverse transcriptase. The DNA residing in chromosomes inside the nucleus, with all the biological information to be transferred to the next generation, is calledgenomic DNA(gDNA). These cookies do not store any personal information. Complementary DNA is often used in gene cloning or as gene probes or in the creation of a cDNA library. RNA is transcribed from only one specific strand of duplex DNA. [7] Additional bead-based methods can be used to isolate specific sub-types of RNA (e.g. The first four of these are the most important, as they are used to construct the molecules that are necessary to make up living cells. If you think the answer is ambiguous, state your case. If the statement is TRUE, explain or give a supporting example. [13] Ribosomal RNA may also be depleted to enrich both mRNA and non-poly-adenylated transcripts such as some non-coding RNA.[14]. Plate the silk moth cDNA library (there is no need to use a genomic silk moth library in this experiment), transfer the phage to a filter and incubate with the 32P-probe under appropriate low-stringency conditions, i.e., reduced temperature and high salt until a "reasonable" number of positive signals result. The best possibility is the span of DNA from the codon for the first Trp residue to the first two nucleotides of the codon for Ile. For each entry in column A, select the appropriate entry in column B. pyrA- strains require a pyrimidine and arginine to grow; fol- strains require folic acid to grow but they are also resistant to aminopterin. 5) 16 pt. Focus on the amino acids with the fewest codons: Met and Trp. B. cDNA contain transcription regulatory sequences found Numerade has step-by-step video solutions, matched directly to more than +2,000 textbooks. Mark D. Adams et al. The result of this second step is the union of the first and second exons, with the intron liberated as a lariat intermediate. For this process cDNA is used over gDNA, since prokaryotes cannot spice out introns contained in gDNA. As part of rigorous quality control, gDNA samples are tested by spectrophotometer and electrophoresis, with concentration determined by UV260 measure and plant concentration determined by pico green measurement. d. All release pyrophosphate as a product. cDNA also does not contain any other gDNA that does not directly code for a protein (referred to as non coding DNA). In some cases, a "substitute AAUAAA" may happen to occur in the transcribed region 3' to the gene, in which case this alternative, "cryptic" polyadenylation site could be used. (f) The E. coli ada repair system is an example of an excision repair pathway. 4) 12 pt. Codons are used to call for specific amino acids to be bonded together to form proteins. bgm protein in E. coli. True. The lesion is removed directly from the damaged base. These elements form the basic building blocks of the major macromolecules of life, including carbohydrates, lipids, nucleic acids and proteins. How would you carry out the cross and score transductants? Kpn 1700 Eco Eco 300 Kpn, *--------------------------------------------> *----------------->, <--------------------------------------------* ------------------. 5b) % cotransduction = # growing only on + folate plate /total pyrA+ X 100. mRNA degrades rapidly after it has completed its purpose. This is displayed in Figure 3.Exitronsare introns that are not spliced out, despite containing no coding sequences. It enhances the affinity and increases the amount of time that holoenzyme (a2bb's) is bound to promoter, i.e. (b) A promoter up mutation--one that binds RNAP better--would likely be a neomorph. In the coordinates of the map, this would be 500 (position of BglII) + 1300 = 1800. A frameshift mutation late in the coding sequences would change the frame of the normal termination codon and translation would continue until it reached a stop codon in the new reading frame. And we have to answer which one is the correct one or which one is the true one. c) The base in the 5' position of the anticodon can often pair with several bases in the "wobble" or 3rd position of the codon (e.g. Retroviruses were discovered to have mechanisms for reverse transcription. This means that they can take RNA chains and produce DNA chains from them. The mutation would prevent cleavage and polyadenylation at the usual site, which could have two different consequences. What type of bonds is being cleaved? 14.3 a) The C terminal peptide. cDNA. This generates a 3' hydroxyl on the nucleotide at the 3' end of the first exon, and effectively takes the intron out of the series of transesterifications by forming a lariat structure. Partial sequences of cDNAs are often obtained as expressed sequence tags. The reconstituted virus can be used to infect host plants to create a new generation of virus. You are cloning an aardvark gene, using a bacterial plasmid as a vector. D. cDNA in a cDNA library (in bacteria) is always single These cookies will be stored in your browser only with your consent. The first child of a couple was born with an autosomal recessive disease causing severe mental retardation. 2003-2022 Chegg Inc. All rights reserved. Now reverse the donor and recipient strains. b) All codons have a U at the second position, hence only hydrophobic amino acids are encoded. Synthesis occurs in a 5' to 3' direction. If so, one can determine which alleles are linked to the disease causing mutation by comparing the micro- or minisatellite alleles present in the affected child and the two parents. If protein were the genetic material, the progeny virus would produce large spots. Is a copy of DNA of a messenger RNA molecule which a result of the reverse transcription. b) Lanes 3-8 tell you that complexes A and B are specific, i.e. A single-base change in GAA to form GTA or in GAG to form GTG could account for the Glu Val replacement in sickle-cell hemoglobin. Support your answer. It should be noted that prokaryotes are not capable of splicing out introns.Exonsare a necessary part of the coding system, being retained after introns are spliced out. Ribosomes assemble around mRNA, creating an amino acid chain to create specific proteins. a) Explain how the Constant Spring allele could have arisen via a single-base substitution or by a frameshift mutation. Deoxyribonucleic acid, (DNA) is the molecule that carries the instructions for all aspects of an organisms functions, from growth, to metabolism, to reproduction. Best study tips and tricks for your exams. BioChain provides access to a comprehensive, well-documented tissue bank containing isolated samples that have been tested for contaminants. Some viruses have RNA and no DNA inside the viral particle. Groups of three nucleotides form the smallest, but most well-defined words in the DNA language. Answer: Option C is correct. Therefore, the rRNA primary transcript would be synthesized in 6500 nucleotides /50 nucleotides per sec = 130 sec, or slightly over 2 min. These are the purines: guanine (G) and adenine (A), and the pyrimidines: thymine (T) and cytosine (C). [3], RNA is transcribed from genomic DNA in host cells and is extracted by first lysing cells then purifying RNA utilizing widely-used methods such as phenol-chloroform, silica column, and bead-based RNA extraction methods. 3. Sequence the silk moth cDNA inserts to identify those similar in sequence to the fly bgm coding sequences. These nucleotides are coupled to a deoxyribose sugar and are able to bind to other deoxyribose sugars via phosphate linkages to form long chains, some of which can be well over 100,000,000 molecules long.
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