การสุ่มหาโปรตีนจับซินทินินในกุ้งกุลาดำ

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CDNA library from the Latex of Hevea brasiliensis

Latex from Hevea brasiliensis contains 30-50% (w/w) of natural rubber (cis-1,4-polyisoprene), the important rawmaterial for many rubber industries. We have constructed a cDNA library from the latex of H. brasiliensis to investigate theexpressed genes and molecular events in the latex. We analyzed 412 expressed sequence tags (ESTs). More than 90% of theEST clones showed homology to previously described sequences in public databases. Functional classification of the ESTsshowed that the largest category were proteins of unknown function (30.1%), 11.4% of ESTs encoded for rubber synthesisrelatedproteins (RS) and 8.5% for defense or stress related proteins (DS). Those with no significant homology to knownsequences (NSH) accounted for 8.7%, primary metabolism (PM) and gene expression and RNA metabolism were 7.8% and6.6%, respectively. Other categories included, protein synthesis-related proteins (6.6%), chromatin and DNA metabolism(CDM 3.9%), energy metabolism (EM 3.4%), cellular transport (CT 3.2%), cell structure (CS 3.2%), signal transduction (ST2.2%), secondary metabolism (SM 1.7%), protein fate (PF 2.2%), and reproductive proteins (RP 0.7%)

The Effect of Chitosan on Organogenesis of Oil Palm Embryo-Derived Callus

Zygotic embryos of oil palm (Elaeis guineensis Jacq. var. tenera) were excised and cultured on MS medium containing 3 mg/l 2, 4-D either with or without 0.05% activated charcoal (AC). Improved growth of embryos was obtained on MS medium supplemented with 0.05% AC. Callus cultures were initiated from embryos, young leaves and roots on MS medium containing 2, 4-D, NAA and 0.05% AC. On these media, two morphologically distinct types of white and yellow compact calluses were produced. Green shoots regenerated after several transfers of the yellow compact calluses from zygotic embryos to MS medium supplemented with 15 mg/l chitosan either with or without 5 mg/l 2, 4-D. Histological sectioning revealed that regenerated shoots originated from a clump of meristematic cells that had dense cytoplasm. Regenerated shoots rooted when transferred to MS medium in the presence of 0.05% AC. Transfer of plantlets to soil was achieved. Callus from young seedling leaves and roots did not regenerate shoots or roots in medium containing 2, 4-D or TDZ, with or without chitosan. This finding shows that chitosan can initiate organogenesis in oil palm callus

Computational identification of Penaeus monodon microRNA genes and their targets

MicroRNAs (miRNAs) are a distinct class of small non-coding RNAs, ~22 nt long, found in a wide variety of organisms.They play important regulatory roles by silencing gene activities at the post-transcriptional level. In this work, we developeda computational workflow to identify conserved miRNA genes in the 10,536 unique Penaeus monodon expressed sequencetags (ESTs). After removing all simple repeats and coding regions in the ESTs, the workflow uses both the conservationof miRNA sequences and several filters obtained from pre-miRNA secondary structure properties to identify conservedmiRNAs. Finally, we discovered six potential conserved miRNA genes such as mir-4152, mir-466k, miR-32*, lin-4, mir-1346 andmir-4310

Evaluation of antibacterial, antioxidant activity and Calmodulin gene expression of Scoparia dulcis Linn.

In Thailand, Scoparia dulcis is being used as a traditional medicine. The extract displays the best inhibitory effect against Pseudomonas aeruginosa and its minimum inhibitory concentration (MIC) was 400 µg/mL followed by Escherichia coli and Staphylococcus aureus with MIC 500 µg/mL. The presence of the extract significantly protected Sf9 cells against H2O2- induced cell death. In addition, the extract also showed DNA damage inhibitory effect in a concentration-dependent manner (0.5- 4 mg/mL). However, at higher concentrations (7.5-30 mg/mL) it might induce damage to the DNA. The prevention of DNA damage differs in different parts of the plant. To support secondary metabolite synthesis in different parts of S. dulcis, we investigated the expression of the Calmodulin gene that is involved in secondary metabolite production. The Calmodulin gene showed the highest expression in the fruit. This finding justifies the use of S. dulcis in the treatment of diseases caused by bacteria and free radicals

Expression and functional analysis of a transgenic cytochrome P450 monooxygenase in Pueraria mirifica

Isoflavonoids are the main compound in White Kwao Krua (Pueraria mirifica), which is an effective folk medicinal plant endemic to Thailand. It has been widely used for improving human physical and treating diseases. There are substances with estrogenic activities have been isolated from P. mirifica, such as puerarin, daidzein and genistein. Isoflavone synthase (IFS) is one of the key enzymes in Leguminous plants to convert liquiritigenin, liquiritigenin C-glucoside and naringenin chalcone to isoflavonoids. The aim of this research was to enhance the production of isoflavonoids by metabolic engineering. Transgenic plants were constructed by introducing P450 gene (EgP450) which is similar to IFS from oil palm (Elaeis guineensis), into P. mirifica by a biolistic method. After the transgenic plants had proved successfully, isoflavonoids of each group plants were determined by HPLC. The contents of daidzein and genistein in transgenic plants were higher than the control plants

Molecular cloning and characterization of the late embryogenesis abundant group 4 (EgLEA4) gene from oil palm (Elaeis guineensis Jacq)

The Late-Embryogenesis Abundant group 4 (LEA4) genes is a group of genes that have been reported to be involved in stress and hormone responses. The completed LEA4 cDNA sequence was first obtained from a set of EST sequences of oil palm (Elaeis guineensis Jacq), named as EgLEA4. The open reading frame is 486 bp in length, encoding a deduced amino acid sequence of 161 residues with a molecular weight of 16.5 kDa and a pI value of about 8.0. Five amino acid motif patterns were found in the EgLEA4 (II, I, III, IV and V) and each had a close identity to similar LEA4 patterns of soybean (64%).Comparison of the nucleotide sequences of the cDNA and the genomic DNA demonstrated that the EgLEA4 gene is composed of 2 exons and 1 intron. The 52 untranslated region shows a putative promoter sequence involved in the transcription process, drought stress and hormone responsive elements. RT-PCR analysis showed that the EgLEA4 gene was only expressed in mesocarp, during the late stages of fruit development. It also had a higher expression in induced drought conditions indicating that the EgLEA4 protein may be involved in plant adaptation and stress (drought) responsive pathway

Role of cytochrome P450 monooxygenase in the bioactivation of aflatoxin B1

In a previous study, the gene EgP450 that encodes the proteins of 505 amino acids was isolated from oil palm. The recombinant protein EgP450 is bound to phenylurea-like herbicides which detoxify the substance. Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus sp., is another toxic compound that is known to cause acute toxic effects and act as a hepatocarcinogenic agent. This study aimed to examine the role of EgP450 enzyme in mycotoxin bioactivation in human mesenchymal stem cells (hMSCs). Docking analysis showed that EgP450 is bound to the group of carcinogens, which includes AFB1, n-(2-fluorenyl) acetamide, n-n-butyl-n-butan-4-ol-nitrosamine, n-nitrosodiethylamine, n-nitrosodiethylamine and n-nitrosodimethylamine. An in vivo aflatoxin toxicity test on hMSCs and AFB1 induces the expression of Bmi-1 which is one of the markers for the development of cancer. The presence of EgP450 at 0.15 μg/mL could reduced the Bmi-1 expression in AFB1 induced cells. Moreover, this protein also showed some antioxidant activity. These results exhibited the enormous potential of EgP450 in the detoxification processes

Identifications of SUMO-1 cDNA and Its Expression Patterns in Pacific White Shrimp Litopeanaeus vannamei

Small ubiquitin-like modifiers (SUMO) work in a similar way as ubiquitin to alter the biological properties of a target protein by conjugation. A shrimp SUMO cDNA named LvSUMO-1 was identified in Litopenaeus vannamei. LvSUMO-1 cDNA contains a coding sequence of 282 nucleotides with untranslated regions of 37 bp at 5'-end and 347 bp at 3'-end, respectively. The deduced 93 amino acids exhibit 83% identity with the Western Honeybee SUMO-1, and more than 65% homologies with human and mouse SUMO-1. LvSUMO-1 mRNA is expressed in most L. vannamei tissues with the highest level in hepatopancrease. The mRNA expression of LvSUMO-1 over development stages in L. Vammamei is distinguished by a low level in nauplius stage and relatively high level in postlarva stage with continuous expression until juvenile stage. The LvSUMO-1 protein and its conjugated proteins are detected in both cytoplasm and nucleus in several tissues. Interestingly, LvSUMO-1 mRNA levels are high in abdominal muscle during the premolt stage, wherein it has significant activities of protein degradation, suggesting its possible role in the regulation of shrimp muscle protein degradation

An EgHd3a-like and its alternatively spliced transcripts in the oil palm (Elaeis guineensis)

Heading date 3a (Hd3a) is an important flowering regulator of short-day plants, and plays a role in several developmental processes. The oil palm is commercially grown for the edible oil derived from its fruit. Flowering and fruit development are key factors for successful production, contributing to productivity of the oil palm fruit. Because little is known about the gene organization of Hd3a in the oil palm, the present study aimed at isolating the Hd3a and its regulatory region. Furthermore, alternatively spliced transcripts were cloned, and their expression in different tissues was also investigated. The structure of the oil palm Hd3a (EgHd3a-like) gene and its promoter were established based on the isolation of EgHd3a-like cDNA and genomic DNA. The promoter analysis revealed that it contains two key regulatory elements, CCAAT boxes and an ARR1 motif, which are the binding sites of Hd3a inducers. Other cis-elements corresponding to flowering and organ development were also represented. Expression of the transcript variants was investigated by RT-PCR in the anther, pistil, mesocarp and leaf. EgHd3a-like was expressed in all tissues tested, which supports its multifaceted roles in several developmental processes. The variants were found in all tested tissue types but at different levels, showing some level of tissuespecificity by variant. Taken together, these results indicate that EgHd3a-like is regulated under various conditions and that transcript variants might play an important role in gene function and regulation