Synthesis of Gene for a True Precursor tRNA,Khorana,Alanyl,Transcription,Tyrosine,Gene Cloning,Synthetic Gene

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Isolation, Sequencing and Synthesis of Genes Isolation, Sequencing and Synthesis of Genes - Introduction Isolation of Genes Isolation of Genes Coding for Specific Proteins Isolation of Genes with Known or Unknown Products having Tissue Specific Expression Isolation of Genes with Known or Unknown Products Using DNA or RNA Probes Use of Heterologus Probes Use of Synthetic Probes Isolation of Genes Coding Use of Transposable Elements Transposon Tagging T-DNA-Insertion Mutagenesis for Isolation of Plant Genes Promoter Enhancer and Gene Trap for Isolation of Genes Differential Screeing and Differential Display Technique for Isolation of Genes Subtractive Hybridizatioon for Gene Isolation Map Based Cloning for Gene Isolation Isolation of Novel Genes by Asis Data at JNU New Delhi

Home >> Biotechnology and Genomics >> Isolation, Sequencing and Synthesis of Genes >> Synthesis of Gene for a True Precursor tRNA

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Synthesis of Gene for a Ttrue Precursor tRNA
Before Khorana could complete the synthesis of gene for yeast alanyl tRNA in 1970, as outlined above, it became obvious that tRNA was not the first direct product of transcription. Instead a precursor molecule is first synthesized which subsequently, after losing segments of RNA by cleavage, give rise to tRNA. Obviously, therefore, the actual gene for yeast alanyl tRNA will be longer than the DNA duplex synthesized by Khorana. In view of this, Khorana subsequently initiated synthesis of a gene for E. coli tyrosine suppressor tRNA precursor DNA duplex which will give rise to this tRNA precursor, was synthesized in the form of 26 small oligonucleotide segments. These were then arranged into six DNA duplex fragments having single stranded ends.

Different parts of complete E. coli tyrosine suppressor tRNA gene

Different Parts of Complete E.coli Tyrosine Suppressor tRNA Gene

These six fragments gave rise to presumed gene for E. coli tyrosine suppressor tRNA precursor. This gene, however, still lacked promoter region and other sequences essential for processing.

Later, in 1979, Khorana reported completion of the total synthesis of a biologically functional tyrosine suppressor transfer RNA gene carrying all regulatory sequences. This gene was 207 base pairs long and included the following:
(i) a 51 base pairs long DNA promoter region;
(ii) a 126 base pairs long DNA corresponding to the precursor tRNA and
(iii) a 25 base pairs long duplex DNA corresponding to 16 base pairs adjoining CCA end of tRNA and the remainder, a modified sequence including EcoRI endonuclease specific sequence.

The details of the complete gene are shown in. The complete synthetic gene was cloned in a vector by one of the gene cloning methods discussed in Chapter 3. On transformation of E. coli, the phage could multiply with the cloned gene.

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Sequencing of a Gene or a DNA Fragment Maxam and Gilbert's Chemical Degradation Method Sanger's Dideoxynucleotide Synthetic Method Automatic DNA Sequencers Slab Gel Versus Capillary Sequencers Slab Gel Sequencing Systems Capillary Gel Sequencing Systems Direct DNA Sequencing Using PCR Ligation Mediated PCR LMPCR DNA Sequencing Through Transcription DNA Sequencing by Hybridization Using Microarrays on DNA Chips DNA Sequencing by DE-MALDI-TOF Mass Spectrometry Synthesis of Genes Synthesis of Gene for Yeast Alanyl tRNA Synthesis of Oligonucleotides Synthesis of Three Duplex Fragments Synthesis of Gene from Three Duplex Fragments Synthesis of Gene for a True Precursor tRNA Total Synthesis of a Human Leukocyte Interferon Gene Gene Synthesis Machines Protection Chemistry and Amidites Permanent Protecting Groups Temporary Protecting Groups Synthesis of Gene Using PCR Synthesis of Genes From mRNA