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MOLECULAR BIOLOGY PART-2 GENE GENE CONCEPT PROTEIN SYNTHESIS LAC OPERON CONCEPT GENE AND GENE CONCEPT To know about the scientists who coined various terminologies and proposed gene hypothesis Gene was coined by W Johansen in 1909 Gene theory was proposed by T H Morgan in 1911 L Pauling P li and d Ingram(1949) I (1949) established the role of genes in protein synthesis The fine structure of gene was proposed by Seymour Benzere(1962) GENE Æ CISTRON Æ RECON Æ MUTON E R Garrod (1908) proposed One g gene – one p product hypothesis G W Beadle and E L Tatum (1948) proposed One gene – one enzyme hypothesis Yanofsky (1965) proposed One gene ( cistron) – one polypeptide hypothesis EUKARYOTIC GENE PROKARYOTIC GENE (Unspilt gene) Has only coding exons C1 E C2 E C3 E (spilt gene) Has only coding exons g introns and non coding Transcription E E E (Polycistronic mRNA) Translation E I E I E Transcription I E E ( Hn – RNA) RNA S Splicing li i I I I E E E E (Monocistronic mRNA) Translation Many polypeptides or proteins t i One protein (spilt gene – Philip A Sharp and Richards Roberts- 1979, Noble prize 1993) CENTRAL DOGMA OF MOLECULAR BIOLOGY PROPOSED BY CRICK (1958) DNA REPLICATION DNA TRANSCRIPTION REVERSE TRANSCRIPTION mRNA TRANSLATION Polypeptide yp p PROTEIN SYNTHESIS Sensestrand 5’.. ATGGCGCACTAA.. 5 ATGGCGCACTAA 3 3’ DNA Antisensestrand 3’..TACCGCGTGATT.. 3 TACCGCGTGATT 5 5’ TRANSCRIPTION mRNA 5’..AUGGCGCAC 5 AUGGCGCAC UAA..3 UAA 3’ TRANSLATION tRNA POLYPEPTIDE 3’..UACCGCGUG…….. 3 UACCGCGUG 5’ 5 aa1- aa2 - aa3 PROTEIN SYNTHESIS IN PROKARYOTE TRANSCRIPTION DNA RNA polymerase Mg++ g mRNA STEPS OF TRANSLATION a)) Activation of amino acid Amino acid t-RNA amino acyl synthetase Mg++, ATP transformylase amino acyl t-RNA (aa tRNA) (aa-tRNA) b) Chain initiation: mRNA IF2 IF3 aa-tRNA aa tRNA GTP Smaller sub-unit of ribosome (30S) IF1 Larger sub sub-unit unit of ribosome (50S) c) Chain elongation: mRNA + ribosome ( 70S) EF – TU EF – TS , GTP peptidyl transferase EF – G ( translocase ) elongating polypeptide chain d) Chain Ch i termination: t i ti Elongating g g polypeptide p yp p chain RF1- recognizes UAA & UAG RF2 recognizes RF2i UAA & UGA RF3- function unknown Peptidyl synthetase Rib Ribosome dissociation di i ti factor f t Nascent p polypeptide yp p chain e) Chain modification: Nascent polypeptide chain Deformylase Exopeptidase Molecular choperones Functional / Structural protein ( having one or more polypeptides) LAC OPERON LAC OPERON - SWITCHED ON LAC OPERON – SWITCHED OFF Q - Which of the following is wrongly matched? 1) cistron – functional unit of gene 2) recon – unit it off recombination bi ti off a gene 3) muton – unit of mutation of a gene 4)) exon – unit of split p gene g which does not carry information Q - The splicing of Hn RNA helps in 1) removal of exons 2) removal of coding segments 3) removal of introns 4) removal of coding exons Q -The hypothesis proposed by G W Beadle and E.L. G.W.Beadle E L Tatum based on their experiment in Neurospora crassa is 1) one gene- one enzyme 2) one gene- one polypeptide 3) one genegene one product 4) one gene- one function E R Garrod(1908) proposed One g gene – one p product hypothesis G W Beadle ead e a and d E L Tatum(1948) atu ( 9 8) proposed One gene – one enzyme hypothesis Yanofsky(1965) proposed One gene ( cistron) – one polypeptide hypothesis Q -The hypothesis proposed by G.W.Beadle and E.L. Tatum based on their experiment in p crassa is Neurospora 1) one gene- one enzyme 2) one gene gene- one polypeptide 3) one gene- one product 4) one gene- one function Q - In Eukaryotes, y , the mRNA is transcribed from DNA with the help of the enzyme 1) RNA primase 2) RNA poly I 3) RNA poly II 4) RNA poly III Q - During transcription mRNA is synthesized in ---- direction and d the th template t l t strand t d off DNA is read in ---- direction and during translation mRNA is read in ---- direction. 1) 5’ →3’, 3’ → 5’ and 5’ → 3’ 2) 5’ 5 →3 →3’, 5 5’ → 3 3’ and 5 5’ → 3 3’ 3) 3’ →5’, 3’ → 5’ and3’ → 5’ 4) 3’ →5’, 5’ → 3’ and 5’ → 3’ Sensestrand PROTEIN SYNTHESIS 5’.. ATGGCGCACTAA.. 3’ DNA Antisensestrand (Template strand) 3’..TACCGCGTGATT.. 5’ mRNA 5’.. AUGGCGCACUAA.. 5 AUGGCGCACUAA 3 3’ TRANSCRIPTION TRANSLATION tRNA POLYPEPTIDE 3’.. UACCGCGUG.. 3 UACCGCGUG 5 5’ aa1—aa2—aa3 aa1 aa2 aa3 Q - During D i transcription t i ti mRNA RNA is i synthesized in ---- direction and the template strand of DNA is read in --- direction and during translation mRNA is read in ---- direction. 1) 5’ 5 →3’, 3,3 3’ → 5 5’ and 5 5’ → 3 3’ 2) 5’ →3’, 5’ → 3’ and 5’ → 3’ 3) 3’ →5’, 5’ 3’ → 5’ and3’ d3’ → 5’ 4)) 3’ →5’,, 5’ → 3’ and 5’ → 3’ Q - What sequence in the template strand of DNA corresponds to the first amino acid inserted i t protein? into t i ? 1) TAC 2) AUG 3) UAG 4) AAA PROTEIN SYNTHESIS Sensestrand 5’.. ATGGCGCACTAA.. 3’ Antisensestrand 3’..TACCGCGTGATT.. 5’ TRANSCRIPTION mRNA 5’.. AUGGCGCACUAA.. 5 AUGGCGCACUAA 3 3’ TRANSLATION tRNA POLYPEPTIDE 3’.. UACCGCGUG.. 3 UACCGCGUG 5’ 5 aa1—aa2—aa3 aa1 aa2 aa3 Q - What sequence in the template strand of DNA corresponds to the first amino acid inserted into protein? 1) TAC 2) AUG 3)) UAG 4) AAA Q - In protein synthesis, synthesis translation involves 1) decoding of triplet codons of tRNA by y anticodons of mRNA 2) decoding of triplet codons of mRNA by anticodons of tRNA 3) decoding of triplet codons of rRNA by anticodons of tRNA 4) synthesis of mRNA from DNA Q - Which of the following statement is wrong with respect to chain initiation step of protein synthesis in E.coli? 1)) 5’ end of mRNA attaches to 30S ribosome with the help of IF3 2)) Formyl y methionine tRNA decodes AUG with the help of IF2 3)) 50S ribosome attaches to the initiation complex with the help of IF1 4)) This p process derives the energy gy from ATP Chain initiation mRNA IF2 IF3 aa-tRNA aa tRNA GTP Smaller sub-unit of ribosome (30S) IF1 (INITIATION COMPLEX) Larger sub-unit of ribosome (50S) Q - Which of the following statement is wrong g with respect p to chain initiation step of protein synthesis in E.coli 1)) 5’ end of mRNA attaches to 30S ribosome with the help of IF3 2) Formyl methionine tRNA decodes AUG with the help of IF2 3) 50S ribosome attaches to the initiation complex l with ith the th h help l off IF1 4) This process derives the energy from ATP Q - Which of the following factors are labelled as translocase that help in the movement of ribosome on mRNA? 1) EF – TU and EF- TS 2) EF- G and eEF2 3) eEF1 and eEF1B 4) 23S and 28S rRNA Q - Peptide bond formation occurs during protein synthesis with the help of the enzyme 1)Amino acyl tRNA synthetase 2)Peptidyl ) p y transferase 3)Peptidyl synthetase 4)Transformylase Q - If a segment of an mRNA has a sequence q 5’ GUACCGAUC 3’ , which of the following could have been the template DNA molecule? 1) 3’CAUGGCUAG 5’ 2) 5’CATGGCTAG3’ 3) 5 5’CAUGGCUAG3’ C UGGCU G3 4) 5’GATCGGTAC3’ Sensestrand 5’..… GTACCGATC….. 3’ (CODOGEN) Antisensestrand (CRYPTOGRAM) 3’..… CATGGCTAG….. 5’ mRNA 5’..… GUACCGAUC….. 5 GUACCGAUC 3’ 3 (CODON) tRNA (ANTICODON) 3’..… CAUGGCUAG….. 5’ Q - If a segment of an mRNA has a sequence 5’ GUACCGAUC 3’ , which of the following could have been the template DNA molecule? 1) 3’CAUGGCUAG 5’ 2) 5’CATGGCTAG3’ 3)) 5’CAUGGCUAG3’ 4) 5’GATCGGTAC3’ Q - A fragment of DNA nucleotide arrangement is 5’ ….ATGGCAGCTTAT….3 5 ATGGCAGCTTAT 3’ What will be the anticodons in tRNAs which will translate the codons p produced by y this part p of DNA? 1) 5’UAC3’ 2) 3’UAC5’ 3 UAC5 3) 5’AUG3’ 4) 3’AUG5’ 5’CGU3’ 3’CGU5’ 3 CGU5 5’GCA3’ 3’GCA5’ 5’CGA3’ 3’CGA5’ 3 CGA5 5’GCU3’ 3’GCU5’ 5’AUA3’ 3’AUA5’ 3 AUA5 5’UAU3’ 3’UAU5’ 5’ ATGGCAGCTTAT 3’ ( Sensestrand ) noncoding Anticodons of tRNA are complementary to sense strand t d (noncoding) ( di ) 3’ UACCGUCGAAUA 5’(tRNA) 1) 5’UAC3’ 2) 3’UAC5’ 3)) 5’AUG3’ 4) 3’AUG5’ 5’CGU3’ 3’CGU5’ 5’GCA3’ 3’GCA5’ 5’CGA3’ 3’CGA5’ 5’GCU3’ 3’GCU5’ 5’AUA3’ 3’AUA5’ 5’UAU3’ 3’UAU5’ Q - The reading frame of mRNA having 1002 nucleotides translated into a nascent polypeptide having amino acids numbering 1) 333 2) 666 3) 999 4) 1001 Q- Consider the statements given below and find the answer among the options given below. Statement A: UAA, UAG and UGA codons d i mRNA in RNA tterminate i t th the synthesis of poylpeptide chain Statement B: UAA, UAG and UGA codons codo sa are e not ot recognized ecog ed by ttRNA 1) Both statements A and B are correct and B is not the reason for A 2) Both statements A and B are correct and B is the reason for A 3) Statement A is correct and B is wrong 4) Statement A wrong and B is correct Q-The replacement of any onenucleotide base in a codon changing the specificity for amino acid is 1)Mis sense mutation 1)Mis-sense 2) non-sense mutation 3) silent mutation 4) frame shift/gibberish mutation Q- Which among the following is wrongly l matched? t h d? 1) Lac Z gene – β galactosidase 2) Lac y gene – β galactoside permease 3) Lac A gene – Transacetylase 4)) Operator p gene g – Repressor p protein p Q- Operator gene of Lac operon i turned is t d on when h lactose l t molecules bind to 1) promoter site 2) operator site 3) regulator gene 4) repressor protein LAC OPERON - SWITCHED ON Q- Operator gene of Lac operon is turned on when lactose molecules bind to 1) promoter t site it 2)) operator site 3) regulator gene 4) repressor protein Q- A synthetic mRNA of repeating sequence 5 CA CA CA CA CA CA CA C…3 5’ C 3’ is used for a cell-free protein synthesizing system like the one used by Nirenberg. If we assume that protein synthesis can begin without the need for an initiator codon what product or products codon, would you expect to occur after protein synthesis? 1) one protein, protein with an alternating sequence of two different types of amino acids 2) one protein, consisting of a single type of amino acid 3) two proteins with an alternating sequence of two different types of amino acids 4) one protein t i with ith an alternating lt ti sequence of three different types of amino acids mRNA 5’ CACACACACACA 3’ (codon) tRNA 3’ GUGUGUGUGUGU 5’ ( (anticodon) ) POLYPEPTIDE Val – Cys – Val – Cys ( i (aminoacids) id ) 1) one protein, protein with an alternating sequence of two different types of amino acids 2) one protein, consisting of a single type of amino acid 3) two proteins with an alternating sequence of two different types of amino acids 4) one protein t i with ith an alternating lt ti sequence of three different types of amino acids Q- Match the following operon genes Column I A)) Structural gene g B) Operator gene C) Promoter gene D) Regulator gene Column II i)) Binding g site for repressor gene ii) Codes for repressor protein iii) Codes for enzyme proteins t i iv) Binding site for RNA polymerase 1) 2) 3) 4) A i ii iv iii B iii i i i C ii iii ii iv D iv iv iii ii Q-The Pribnow box is represented by 1) 2) 3) 4) 5` ATATTA 3` 5` 5 AATAAT 3 3` 5` TATAAT 3` 5` TATAA 3`