Cdna Cloning And Sequence Analysis Essay - 1116 Words

3. Results 3.1 cDNA cloning and sequence analysis In this study, we cloned the rabbit NMU, NUM-R1 and NMU-R2 cDNA fragments using RT-PCR. Specific fragments were amplified and target gene fragment length was 443, 366 and 203 bp, respectively (Fig.1 A, B and C), consistent with the expected size of the fragment, and clear bands. These fragments were confirmed by sequencing, and the NMU, NMU-R1 and NMU-R2 sequences were submitted to GenBank (GenBank IDs: KP276160.1, KM-236787.1 and KM-236788.1). NCBI database map views program analysis showed that The rabbit NMU, NMU-R1 and NMU-R2 gene were located at chromosome 2, 3 and 1 respectively. The sequence analysis showed that rabbit NMU gene exhibited an open reading frame (ORF) of 564 nucleotides encoding 188 amino acids residues, while NMU-R1 gene included 360 nucleotides encoding 120 amino acids residues, and NMU-R2 gene included 288 nucleotides encoding 96 amino acids residues. The rabbit NMU, NMU-R1 and NMU-R2 genes sequences were 84%, 84% and 87% identical to those of the corresponding human homologues, respectively. The homology of the nucleotide and amino acid sequences are compared in Table (2, 3 and 4). Multiple alignment analysis of the amino acid sequences indicated that the rabbit NMU, NMU-R1 and NMU-R2 were relatively conserved in human, mouse, cow, pig, rat and sheep (Fig. 2, 3 and 4). Phylogenetic analysis revealed that the NMU coding sequences are highly conserved between the related species (Fig. 5). PhylogeneticShow MoreRelatedBiological Molecules Like Nucleic Acids And Polysachharides2245 Words   |  9 Pagesstates. To know the human genome, itâ€⠄¢s really need to understand the proteome of humans. It has been concluded that from the point of view of therapeutics that the targets for major drugs are proteins instead of nucleic acids (Verrills, 2006). Protein analysis identifies changes like its expressions, protein-protein interactions, post-translational modifications and sub-cellular distribution. Nowadays many different molecular techniques are being used as tools to discover and determine different biologicalRead MoreApple Scar Skin Viroid Case Study1156 Words   |  5 Pagesimprove the LAMP selectivity and promote its diagnostic use in detecting ASSVd in plants. ASSVd LAMP was accelerated by the use of loop primers and highly sensitive with a detection limit of 103 copies of ASSVd cDNA per reaction within 30 min. The real-time LAMP and melting curve analysis could differentiate between the true-positive LAMP amplicons and false-positive non-specific primer amplification products. The optimized RT-LAMP was then followe d by the addition of AuNP:polyA10-ASSVd LAMP barcodesRead MoreSugarcane ( Saccharum Spp )1158 Words   |  5 Pages2008; Gui et al. 2011), and woody plants (Allina et al. 1998; Hu et al. 1998; Harding et al. 2002) but in sugarcane it has not been cloned and charaterised till now. Therefore in this present study, we have attempted and cloned the partial coding sequences of 4CL from EC11010, the energy cane and characterized to understand the regulatory reactions of lignin biosynthesis pathway in sugarcane which ultimately can be used for delignification of sugarcane biomass and its improved utilization for ethanolRead MoreBased on the Plant Biology Journal entitled â€Å"Gene discovery in the wood-forming tissues of poplar:900 Words   |  4 PagesBased on the Plant Biology Journal entitled â€Å"Gene discovery in the wood-forming tissues of poplar: Analysis of 5,692 expressed sequence tags†, by Sterky et al., 1998, expressed sequence tags (EST) database was prepared with s pecific procedure. First of all, EST can be defined as a short fragments sequence, range from 100 bp- 1000 bp. It derived from a c DNA clone which represents the expressed genes sequence. Usually, ESTs are used to identify the full length genes and serves as a target for mapping withRead MoreA Research Study On Cultured And Handled At 20. C Using Standard Methods ( Brenner ) Essay1268 Words   |  6 Pagesinterference RNAi depletion of cdc-25.2 in mutants of E3 ubiquitin-ligase complex genes was performed by the soaking RNAi method as previously described (Lee et al., 2016). The cdc-25.2 dsRNA was prepared by in vitro transcription using a cdc-25.2 cDNA clone, yk472b2, as the DNA template. Worms synchronized at the first larval (L1) stage were soaked into the dsRNA solution and incubated for 48 h at 20  °C. Then, the soaked animals were transferred to OP50-seeded NGM plates and allowed to grow untilRead MoreThe Viological Structure Of G-Protein Coupled Receptorss1463 Words   |  6 Pageseffectors (Lefkowitz, 2007). EXPERIMENT 2: Cloning of the gene and cDNA for mammalian ÃŽ ²-adrenergic receptor and homology with rhodopsin (Dixon RA, et al., 1986) Overview of the Experiment The ligand binding technique mentioned in experiment 1 was also used to tag and isolate receptor molecules through various stages of purification. After successfully purifying ÃŽ ²AR in solubilised form using digitonin (Lefkowitz, 2007), it was then used to clone ÃŽ ²2AR cDNA from hamsters. Methodology Constructing degenerateRead MoreExpressed Sequence Tags (ESTs) versus Serial Analysis of Gene Expression800 Words   |  4 PagesExpressed Sequence Tags (ESTs) versus Serial Analysis of Gene Expression (SAGE) INTRODUCTION Expressed sequence tag or also known as EST are 200 to 800 unedited nucleotide bases in length, and randomly selected single pass sequence reads which are derived from the cDNA libraries while serial analysis of gene expression or SAGE have more shorter sequence tags with only 10 to 20 base pairs. But even it is short it still have enough information to uniquely identify a transcript, especially if it isRead MoreNucleic Acid Measurements1302 Words   |  6 Pagessulfate reduction – nirS (nitrate reductase) nitrate reduction Wednesday, July 24, 2013 16S rRNA gene probes †¢ Oligonucleotide primers for PCR Target region cDNA 16S rDNA clone library †¢ Oligonucleotide probes complementary to 16S rRNA molecule – no need for PCR because many copies in cells Wednesday, July 24, 2013 cDNA RNA ribosome Secondary Structure: 16S rRNA Different locations on the 16S rRNA molecule offer identity at different phylogenetic levels †¢Domain EU338 †¢PhylumRead MoreThe Structural Domains And Mechanism Of Action Of Ribozymes1686 Words   |  7 Pagesconsist of a catalytic domain and a substrate binding domain. (Ebrahimian, 2015) The substrate binding domain has a specific sequence antisense to the target mRNA. This sequence recognizes and hybridizes specifically to its substrate. (Missailidis, 2008). Alteration of the substrate binding domain can be done so that the substrate specifically cleaves any mRNA sequence. The RNA catalytic domain cleaves the substrate at a target site recognized by the ribozyme (Glick Pasternak, 2003). The resultantRead MoreBiology Essay3361 Words   |  14 PagesSkill: Factual Recall 4) The production of multiple identical copies of gene-sized pieces of DNA defines A) gene cloning. B) plasmid transformation. C) clonal selection. D) tissue culturing. E) plasmolysis. Answer: A Topic: 12.1 Skill: Factual Recall 5) In the process of human gene cloning using recombinant plasmids, the bacterial plasmid A) functions as a vector. B) is the source of the gene