Insights from the cDNA and EST analysis of Antrodia cinnamomea

It is of interest to document the insights gleaned from the cDNA and EST analysis of Antrodia cinnamomea (a fungal species). Hence a library of sequences was constructed and analysed using standard procedures to gain new insights. Therefore, 65 ESTs, with size ranging from 300-2000 bp, were constructed. This included 46 ESTs with definite annotation, 18 ESTs were hypothetical and 1 new protein derived from BLAST analysis. We assigned 227 Gene Ontology terms linked to cell composition, transport, catalytic activity, and regulation functions in these sequences. Moreover, 56 matching genes were found in 8 Kyoto Encyclopedia of Genes and Genomes pathways. Data also showed 271 SSRs from Antrodia cinnamomea ESTs with an occurrence frequency of 96.82%. The STRING data analysis showed 29 genes encoded enzymes highly involved in protein-to-protein interactions linked to expression of regulation function. Thus, we documented some insights from the cDNA and EST analysis of Antrodia cinnamomea for further data mining.

Molecular Cloning, Structure and Phylogenetic Analysis of a Hemocyanin Subunit from the Black Sea Crustacean Eriphia verrucosa (Crustacea, Malacostraca)

Hemocyanins are copper-binding proteins that play a crucial role in the physiological processes in crustaceans. In this study, the cDNA encoding hemocyanin subunit 5 from the Black sea crab Eriphia verrucosa (EvHc5) was cloned using EST analysis, RT-PCR and rapid amplification of the cDNA ends (RACE) approach. The full-length cDNA of EvHc5 was 2254 bp, consisting of a 5′ and 3′ untranslated regions and an open reading frame of 2022 bp, encoding a protein consisting of 674 amino acid residues. The protein has an N-terminal signal peptide of 14 amino acids as is expected for proteins synthesized in hepatopancreas tubule cells and secreted into the hemolymph. The 3D model showed the presence of three functional domains and six conserved histidine residues that participate in the formation of the copper active site in Domain 2. The EvHc5 is O-glycosylated and the glycan is exposed on the surface of the subunit similar to Panulirus interruptus. The phylogenetic analysis has shown its close grouping with γ-type of hemocyanins of other crustacean species belonging to order Decapoda, infraorder Brachyura.

PENGARUH TERAPI AKTIVITAS KELOMPOK SOSIALISASI TERHADAP KEMAMPUAN SOSIALISASI KLIEN ISOLASI SOSIAL

<p><em>Social isolation is a diagnosis 3 largest of the diagnosis inthe area of jambi province. This is given to the isolation of social one is thheraupeutic activity of socialization. Social isolation is a disorder of interpersonal relationships that occur due to the inflexible personality, giving rise to maladaptive behavior and interfere with the functioning of a person in touch. Socialization Activity Group Therapy is seeking to facilitate the socialization of a number of clients with impaired social relationships in groups. The purpose of study was to determine the influence of group activity socialization toward among clients with impaired social. This study design using the one group pretest-posttest, the sampling technique is purposive sampling of 12 respondents. Client socialization skills were measured before and after intervention using observation sheet Est. Analysis of data with paired samples T-test. The analysis shows value of the average capability of the respondents before TAKS 2,42 and after TAKS 19,00.The analysis shows a significant influence on the taks on socialization skills with p = 0.009. It is concluded that there is influence of group activity therapy disseminate the client socialization ability of social isolation in inpatient psychiatric hospitals of the province of Jambi in 2016.</em></p><p><em><br /></em></p><p><em><br /></em>Isolasi sosial merupakan diagnosa 3 terbesar dari 7 diagnosa yang ada di rumah sakit jiwa daerah provinsi Jambi. Terapi yang diberikan kepada klien isolasi sosial salah satunya adalah terapi aktivitas kelompok sosialisasi. Isolasi sosial adalah gangguan hubungan interpersonal yang terjadi akibat adanya kepribadian yang tidak fleksibel, sehingga menimbulkan prilaku yang maladaptif dan mengganggu fungsi seseorang dalam berhubungan. Terapi Aktivitas Kelompok Sosialisasi (TAKS) adalah upaya memfasilitasi sosialisasi sejumlah klien dengan kerusakan hubungan social secara kelompok. Tujuan dari penelitian ini untuk mengetahui pengaruh TAKS terhadap kemampuan sosialisasi klien isolasi sosial.Desain penelitian ini menggunakan rancangan <em>the one group pretest-postest</em>, dengan teknik pengambilan sampel secara purposive sampling terhadap 12 responden. Kemampuan sosialisasi klien diukur sebelum dan sesudah dilakukan intervensi TAKS menggunakan lembar observasi. Hasil analisa data menunjukkan nilai rata-rata kemampuan sosialisasi responden sebelum diberikan TAKS adalah 2,42 dan sesudah di berikan TAKS menunjukan nilai rata-rata 19,00. Analisa data dengan uji paired sample T-test menunjukkan adanya pengaruh yang signifikan dari TAKS terhadap kemampuan sosialisasi dengan p=0,009. Penelitian ini ada pengaruh terapi aktivitas kelompok sosialisasi terhadap kemampuan sosialisasi klien isolasi sosial di ruang rawat inap rumah sakit jiwa daerah provinsi Jambi tahun 2016.</p>

Description of a Zostera marina catalase gene involved in responses to temperature stress

Catalase (CAT) is an antioxidant enzyme that plays a significant role in cellular protection against oxidative damage by degradation of hydrogen peroxide to oxygen and water. In the present study, the complete CAT cDNA sequence of Zostera marina was identified through expressed sequence tags (EST) analysis and the rapid amplification of cDNA ends (RACE) technique. The nucleotide sequence of ZmCAT cDNA consisted of 1,816 bp with a 1,434 bp open reading frame (ORF), encoding a polypeptide of 477 amino acid residues, which possessed significant homology to other known plant CATs. The molecular mass of the predicted protein was 55.3 kDa with an estimated isoelectric point of 6.40. Phylogenetic analysis showed that ZmCAT was closely related to CAT from gramineous species. In response to temperature stress, H2O2 and MDA contents in Z. marina increased significantly with cold stress (<10 °C) and heat stress (>25 °C). ZmCAT expression was significantly upregulated at temperatures from 5 to 10 °C and then gradually downregulated, reaching its lowest expression at 30 °C. Recombinant ZmCAT protein exhibited strong antioxidant activity over a wide temperature range, with the highest rZmCAT activity observed at 25 °C and a higher relative activity retained even with heat stress. All these results indicated that ZmCAT was a member of the plant CAT family and involved in minimizing oxidative damage effects in Z. marina under temperature stress.

Total RNA Isolation and cDNA synthesis from Bixa orellana bark

<p><strong>Annatto is one of the important natural food colourant widely used in dairy industry. Quality RNA in large quantity is often required in the analysis of gene expression. RNA extraction from samples collected from woody plants is generally complex, and becomes the main limitation to study gene expression, particularly in crops like <em>Bixa orellana</em>. Standard RNA extraction protocols are time consuming, laborious and cannot be adapted for high throughput functional analysis. Therefore a simple and effective protocol for extraction of high quality total RNA from bark tissue of woody stem was achieved using the RNeasy plant mini kit (Qiagen, USA). The extracted RNA was successfully converted into double-stranded cDNA using the SMATer cDNA synthesis kit (Clontech, USA) which is based on the <span style="text-decoration: underline;">S</span>witching <span style="text-decoration: underline;">M</span>echanism <span style="text-decoration: underline;">A</span>t 5’ End of <span style="text-decoration: underline;">R</span>NA <span style="text-decoration: underline;">T</span>ranscript (SMART) technology. The integrity of the total RNA used for synthesizing double stranded cDNA was assessed agarose gel electrophoresis and by PCR. As expected, the PCR product contained the full coding sequence plus 69 and 196 bp of 5’ and 3’ UTRs respectively. The double-stranded cDNA was used successfully for creating a SSH cDNA library. The cDNA could also be useful for a number of other applications like cDNA library construction, EST analysis, RACE and Next Generation Sequencing (NGS).</strong></p>

Construction of cDNA library from Prunus campanulata leaves and preliminary expressed sequence tag (EST) analysis during cold stress

The molecular mechanisms underlying cold-resistance in Prunus campanulata Maxim. (P. campanulata) are not fully understood. This study aimed to establish a full-length library and analyze expressed sequence tags (ESTs) to provide tools to investigate the mechanisms of P. campanulata growth at low temperatures. Based on the switching mechanism at 5’end of RNA transcript technology, a full-1ength cDNA library was generated from young leaves of P. campanulata after 72 h treatment at 1◦C, and a preliminary EST analysis was carried out. Quantitative reverse transcription polymerase chain reaction was used to assess the expression of selected cold-related genes. The cDNA library titer was 1.2 × 106 cfu/mL−1, with a recombinant rate of 96%. The average size of inserted cDNA fragments was 1.3 Kb. EST data revealed the existence of 834 clones representing a total of 667 unigenes, including 574 singletons and 93 contigs. Blast analysis identified 475 unigenes with known and putative functions. Based on similarity search and GO annotation, 84 unigenes were associated with “response to stimuli”, suggesting that cold stress induced significant alterations in gene expression in P. campanulata cultivated at 1◦C for 72 h. Interestingly, DRP, MYB, HSP, GPX and GA20-ox gene expression was significantly up-regulated in plants cultivated at low temperature, while transcript levels of TIL, CDPK were decreased. P. campanulata cultivating at low temperature express genes associated with “response to stimuli”, and in particular DRP, MYB, HSP, GPX and GA20-ox gene are up-regulated while TIL, CDPK are downregulated in response to low temperature-stress