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1.
Template-directed synthesis on the oligonucleotide d(C7-G-C7) 总被引:1,自引:0,他引:1
When the deoxynucleotide template d(C7-G-C7) is incubated with the activated nucleotides 2-MeImpG and 2-MeImpC, a series of oligomers of G up to the sevenmer and a series of copolymers of composition GnC with n = 3 to 13 are formed. Oligomers GnC with n greater than 7 are completely degraded by pancreatic ribonuclease, establishing that they contain a 3' to 5' internucleotide bond between 5'-C and 3'-G within a sequence of the form (pG)ipC(pG)j. As expected, (pG)7-Cp and (pG)6-Cp are major hydrolysis products. Detailed analysis of the product distribution shows that a substantial fraction of the oligomeric products are of the type (pG)ipC(pG)j with i less than 7. This shows that product synthesis does not necessarily begin at the 3' terminus of the template. The significance of this finding in terms of the origin of molecular replication is discussed. 相似文献
2.
The replacement of reporter groups, such as fluorescent molecules or enzymes, by an amplifiable reporter should lead to bioassays of greatly increased sensitivity, since a very large number of copies of the reporter can be accumulated in a short time. Midivariant RNA is an appropriate reporter, since it is autocatalytically replicated by Q beta RNA polymerase in vitro. This RNA can be amplified exponentially, with a population doubling time of 36 seconds, resulting in the synthesis of 10(6) copies of each molecule in 12 minutes. We have used chemical methods to attach biotin to the 5' terminus of midivariant RNA via a disulfide linker. This biotinylated RNA combines with avidin to give a product that is readily purified by gel electrophoresis. The RNA-biotin-avidin adduct, and the RNA released from it by reductive cleavage of the linker arm, replicate normally. The RNA-biotin-avidin adduct should be a suitable reporter for a variety of replication-assisted bioassays involving biotinylated antibodies or biotinylated nucleic acid probes. 相似文献
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6.
Summary The condensation reactions of activated nucleotides, ImpN or 2-MeImpN, with the selfcomplementary ribo-octanucleotide GCGCGCGC or with the partially self-complementary heptanucleotide GCGCGCG were studied. The templatedirected reaction of 2-meImpC with the heptamer yields the 3–5 octamer as the main product. All other reactions yield 2–5-linked octamers and pyrophosphates as major products. Surprisingly, 2-MeImpG facilitates the reaction of 2-MeImpC with the heptamer.Procedures for the analysis by gel electrophoresis of the oligomeric products obtained in reactions of this kind are described. 相似文献
7.
The origin of polynucleotide-directed protein synthesis 总被引:7,自引:0,他引:7
Leslie E. Orgel 《Journal of molecular evolution》1989,29(6):465-474
Summary If protein synthesis evolved in an RNA world it was probably preceded by simpler processes by means of which interaction with
amino acids conferred selective advantage on replicating RNA molecules. It is suggested that, at first, the simple attachment
of amino acids to the 2′(3′)-termini of RNA templates favored initiation of replication at the end of the template rather
than at internal positions. The second stage in the evolution of protein synthesis would probably have been the association
of pairs of charged RNA adaptors in such a way as to favor noncoded formation of peptides. Only after this process had become
efficient could coded synthesis have begun. 相似文献
8.
5-Deoxy-5-nucleosideacetic acids II–V are isostructural analogues of nucleotides with a carboxylate group in the place of the 5-phosphate group. We have studied their oligomerization in aqueous solution using a water-soluble carbodiimide as the condensing agent in the presence or absence of an appropriate polynucleotide template. Condensation of adenylic acid analogues IIa, IIIa, and Va in the presence of polyuridylic acid were found to be the most efficient reactions. Cyclization of the activated monomers to lactones and the insolubility of the oligomers in aqueous solution were found to be obstacles to the efficient formation of long oligomers. 相似文献
9.
Summary We have studied the reactions between adenosine 5-phosphorimidazolide and 9-(2-amino-2-deoxyxylofuranosyl) adenine (I) or 3-methylamino-3-deoxyadenosine (II), both with and without a poly (U) template. We find that both amino compounds react much more rapidly than does adenosine, in the absence of a template. The rate of reaction is greatly enhanced by a poly (U) template in the case of I, but the enhancement is slight in the case of II.Abbreviations A
adenosine
- xylo ANH2
9-(2-amino-2-deoxy--D-xylofuranosyl) adenine
- ANHMe
3-methylamino-3-deoxyadenosine
- ImpA
adenosine 5-phosphorimidazolide
- A3 pA
adenylyl-[35]-adenosine
- A2 pA
adenylyl-[25]-adenosine
- UNPA
adenylyl-[52]-2-amino-2-deoxyuridine
- xylo ANPA
9-[adenylyl-(52)-2-amino-2-deoxy--D-xylofuranosyl]adenine
- A(NMe)pA
adenylyl-[53]-3-methylamino-3-deoxyadenosine
- pA
adenosine 5phosphate
- AppA
P1, P2-diadenosine 5pyrophosphate
- (pA)n
n = 2, 3 [2-5]-linked oligomers of pA
- A2 pA2 pA
[2-5]-linked trinucleoside diphosphate of A
- poly (U)
polyuridylic acid 相似文献
10.
Summary The yields of dipeptide obtained from the reaction of 0.2M 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) and 0.2M amino acid at pH 8.2 ranged from 0.1% to 35.5% for a group of 15 amino acids. The yields of glyser (35.3%), gly-cys (11.8%) and gly-thr (5.4%) were considerably greater than dipeptide yields obtained from any of the other 12 amino acids ( 1.7%). Aminolysis of 0.05M 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) by 0.4M serine ethyl ester yielded 53% glycylserine diketopiperazine, via N-(glycyl)-serine ethyl ester as a transient intermediate. The prebiotic significance of these reactions is discussed.Abbreviations
MepA
adenosine-5-(O-methylphosphate)
-
MepA-gly
2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate)
-
DKP
diketopiperazine
-
serOEt
serine ethyl ester
-
gly-serOEt
N-(glycyl)-serine ethyl ester
-
Boc-gly
N-tertbutyloxycarbonylglycine
-
cyclo-(gly-ser-)
glycylserine diketo-piperazine
-
O-gly-ser
O-glycylserine
-
O-(gly)-gly-ser
O-(glycyl)-glycylserine
-
gly-ser
N-glycylserine 相似文献