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Molecular characteristics of native and recombinant amylolytic biocatalysts of the extreme thermophile Geobacillus thermoleovorans

Sun, 01 Jan 2017 00:00:00 GMT

Title: Molecular characteristics of native and recombinant amylolytic biocatalysts of the extreme thermophile Geobacillus thermoleovorans Authors: Mehta, Deepika; Satyanarayana, Tulasi Abstract: The annual sale of amylolytic enzymes is estimated to reach $2,238.4 million by 2018. Starch hydrolyzing enzymes find major applications in baking, alcohol, detergent and textile industries. Geobacillus thermoleovorans, an extremely thermophilic bacterium, is a well known producer of starch hydrolyzing enzymes. With the advent of high throughput genome sequencing technologies and availability of large amount of sequence data, it has become easy to produce thermostable recombinant enzymes in mesophilic hosts with no loss of enzyme characteristics, making the overall process economical. This review focuses on the native and recombinant starch hydrolyzing enzymes from G. thermoleovorans, their production, characteristics, structure-function aspects and potential industrial applications. Page(s): 9-21

Revisited immune reactivity between native semi-purified protoplasmic (caprine) versus commercial purified protoplasmic (bovine) antigens for the screening of goatherds endemic for Johne’s disease

Sun, 01 Jan 2017 00:00:00 GMT

Title: Revisited immune reactivity between native semi-purified protoplasmic (caprine) versus commercial purified protoplasmic (bovine) antigens for the screening of goatherds endemic for Johne’s disease Authors: Gupta, Saurabh; Singh, Shoor Vir; Bhatia, AK Abstract: The present study compared the immune-reactivity of 3 antigens of Mycobacterium avium subspecies paratuberculosis (MAP) sourced from different geographical regions and livestock species for the diagnosis of Johne’s disease (JD) in goats. Screening of 360 faecal and serum samplesof goats by microscopy, i-ELISA, b-ELISA and r-ELISA gave 56.9, 40.0, 34.4 and5.2% positive samples, respectively. Considering all the 4 tests (microscopy, i-ELISA, b-ELISA & r-ELISA kits), 3.0 and 35.2% goats were found common positive and negative, respectively. Of 3 ELISA tests, i-ELISA had the highest sensitivity, followed by b-ELISA and r-ELISA kit. ‘i-ELISA’ based on ‘indigenous antigen’ recovered from native (‘Indian Bison Type’) MAP strain of goat origin had superior immune reactivity as compared to imported commercial PPAs(protoplasmic antigens) of bovine origin (b-ELISA from Allied Monitor Inc., USA) and commercial ELISA kit for ruminant species (ID Vet, France). Lower immune-reactivity of commercial antigens as compared to ‘indigenous antigen’ indicated that search for universally acceptable ‘ELISA kit’ is not practically possible. Page(s): 22-29

Development and immunereactivity evaluation of a chimeric recombinant protein encoding Brucella antigen: In silico to in vitro

Sun, 01 Jan 2017 00:00:00 GMT

Title: Development and immunereactivity evaluation of a chimeric recombinant protein encoding Brucella antigen: In silico to in vitro Authors: Abdollahi, Abbas; Mansouri, Shahla; Amani, Jafar; Fasihi-Ramandi, Mahdi; Moradi, Mohammad Abstract: Brucellosis is an important health problem in developing countries and no vaccine is available for the prevention of infection in humans. Because of clinically infectious disease and its economic consequences in human and animals, designing a proper vaccine against Brucella is desirable. In the present study, we evaluated the immune responses induced by a designed recombinant chimera protein and investigated the immunogenic potential of some immune reactive antigens of Brucella. Three immune dominant antigens of Brucella including trigger factor (TF), Omp31 and Bp26 (have been characterized as potential immunogenic and protective antigens) were fused together by EAAAK linkers to produce a chimera. Recombinant chimeric protein was synthesized, cloned and expressed in Escherichia coli BL21 (structure were designed in silico). The purification of recombinant protein was performed by using Ni-NTA agarose, and anti-His antibody was used for confirmation (Western blot). The recombinant chimeric protein could be a new potential antigen candidate for the development of a subunit vaccine against Brucella. These results demonstrate the role of the Bioinformatics in vaccine design, assisted by experimental procedures. Page(s): 30-36

Crustacean hyperglycemic hormone family gene silencing in Penaeus monodon mediated through dsRNA synthesized in vitro from genomic and cDNA

Sun, 01 Jan 2017 00:00:00 GMT

Title: Crustacean hyperglycemic hormone family gene silencing in Penaeus monodon mediated through dsRNA synthesized in vitro from genomic and cDNA Authors: Vrinda, S; Reshmi, C; Jose, Seena; Reynold, P; Vijayan, K K; Philip, Rosamma; Singh, I S Bright Abstract: RNA interference (RNAi) is the phenomenon in which long dsRNA is able to silence cognate gene expression. In the present study, 801 bp crustacean hyperglycemic hormone 1 (CHH1) and 795 bp moult inhibiting hormone 1 (MIH1) specific dsRNAs from genomic DNA, and 316 bp gonad inhibiting hormone (GIH) specific dsRNA from cDNA were constructed in vitro. Then the specific dsRNA constructs were administered into adult shrimps (Penaeus monodon). The gene expression was studied by semi quantitative RT-PCR and by monitoring haemolymph glucose concentration, duration of moulting and expression of vitellogenin as measures of specific biological activity. The gene silencing of CHH1, MIH1 and GIH genes could be attained within 24 h of dsRNA application. GIH gene silencing was observed up to 60^th h. However, a complete silencing of MIH1 and MIH2 continued to 108h post administration. Physiology of the animals injected with dsRNA of CHH1, MIH1 and GIH corroborated with the silencing of the specific genes resulting in the decrease of haemolymph glucose level, reduction in the days of moulting and expression of vitellogenin gene, respectively in adult shrimp. These results suggest the possibility of using dsRNAs of CHH family hormone genes as molecular tools for silencing inhibitory genes in turn affecting induced maturation in P. monodon. Page(s): 37-43