Expression analysis of the fatty acid desaturase 2-4 and 2-3 genes from Gossypium hirsutum in transformed yeast cells and transgenic Arabidopsis plants.

Fatty acid desaturase 2 (FAD2) enzymes are phosphatidylcholine desaturases occurring as integral membrane proteins in the endoplasmic reticulum membrane and convert monounsaturated oleic acid into polyunsaturated linoleic acid. The major objective of this research was to study the expression and function of two cotton FAD2 genes (the FAD2-3 and FAD2-4 genes) and their possible role in plant sensitivity to environmental stress, since plants may increase the polyunsaturated phospholipids in membranes under environmental stress events, such as low temperature and osmotic stress. Two FAD2 cDNA clones corresponding to the two FAD2 genes have been isolated from a cotton cDNA library, indicating both genes are truly expressed in cotton. Model yeast cells transformed with two cotton FAD2 genes were used to study the chilling sensitivity, ethanol tolerance, and growth rate of yeast cells. The expression patterns of the two FAD2 genes were analyzed by reverse transcription polymerase chain reactions (RT-PCR) and Western blot analyses in cotton plants under different treatment conditions. The coding regions of both FAD2 genes were inserted downstream from the CaMV 35S promoter in the pMDC gateway binary vector system. Five different FAD2/pMDC constructs were transformed into the Arabidopsis fad2 knockout mutant background, and multiple potential transgenic Arabidopsis plant lines harboring the cotton FAD2 genes were generated. The cotton FAD2 genes were amplified by the polymerase chain reaction (PCR) from the genomic DNAs isolated from the transgenic Arabidopsis T1 plant lines. Complementation of the putative transgenic Arabidopsis plants with the two cotton FAD2 genes was demonstrated by gas chromatography analyses of the fatty acid profiles of leaf tissues. The cellular localization of cotton FAD2-4 polypeptides with N-terminal green fluorescence protein (GFP) was visualized by confocal fluorescence microscopy. The phenotype of transgenic Arabidopsis plants transformed with the cotton FAD2-4 gene was compared to Arabidopsis knockout fad2 mutant plants and wild type Arabidopsis plants regarding their sensitivity to low temperature, and the size and height of the plants

Cluster M Mycobacteriophages Bongo, PegLeg, and Rey with Unusually Large Repertoires of tRNA Isotopes

Genomic analysis of a large set of phages infecting the common hostMycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The cluster M mycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to 83.7 kbp). They exhibit a variety of features not previously described in other mycobacteriophages, including noncanonical genome architectures and several unusual sets of conserved repeated sequences suggesting novel regulatory systems for both transcription and translation. In addition to containing transfer-messenger RNA and RtcB-like RNA ligase genes, their genomes encode 21 to 24 tRNA genes encompassing complete or nearly complete sets of isotypes. We predict that these tRNAs are used in late lytic growth, likely compensating for the degradation or inadequacy of host tRNAs. They may represent a complete set of tRNAs necessary for late lytic growth, especially when taken together with the apparent lack of codons in the same late genes that correspond to tRNAs that the genomes of the phages do not obviously encode

Webinar 5: Teaching Biotech from a Distance - Examples and Sharing

This webinar is part of the Spring 2020 Teaching Biotech from a Distance webinar series provided InnovATEBio National Biotechnology Education Center. In this presentation, Dr. Daiyuan "Daisy" Zhang, Del Mar College, describes how she adjusted her biotechnology teaching strategies when the Coronavirus pandemic forced the college to shut down. This webinar features three examples of lab activities related to SARS-CoV-2 that students completed online. The featured labs covered topics like virus mutations, protein sequencing, and experimental design. A link is provided to Teaching Biotech from a Distance blog and the resources mentioned in the webinar. This webinar runs 1:18:06 minutes in length. Additional webinars in the series are available to view separately.Â

Developing a Highly Capable Biomanufacturing Technical Workforce in South Texas

In this webinar, published by InnovATEBIO, J. Robert Hatherill and Daiyuan Zhang discuss Del Mar College's biomanufacturing ATE project. Del Mar College's biotechnology program, a biopharmaceutical and vaccine manufacturing training program, biotechnology degree requirements, internship outcomes, student assessment data, other biomanufacturing areas for hands-on training, and industry partner recommendations are highlighted. The video recording runs 34:22 minutes in length

ITS2 rDNA VARIATION OF TWO BLACK WIDOW SPECIES, LATRODECTUS MACTANS AND LATRODECTUS HESPERUS (ARANEAE, THERIDIIDAE)

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PKR Protects the Major Catalytic Subunit of PKA Cpk1 from FgBlm10-Mediated Proteasome Degradation in <i>Fusarium graminearum</i>

For optimal proteolytic function, the proteasome core (CP or 20S) must associate with activators. The cAMP-PKA pathway is reported to affect the activity of the proteasome in humans. However, the relationship between the proteasome and PKA is not well characterized. Our results showed that the major catalytic subunit Cpk1 was degraded without the protection of Pkr. Eleven (out of 67) pkr suppressors had FgBlm10 C-terminal truncation, one suppressor had an amino acid change mutation in the PRE6 ortholog (FGRRES_07282), and one in the PRE5 ortholog (FGRRES_05222). These mutations rescued the defects in growth and conidial morphology, Cpk1 stability, and PKA activities in the pkr mutant. The interaction of FgBlm10 with FgPre5 and FgPre6 were detected by co-immunoprecipitation, and the essential elements for their interaction were characterized, including the FgBlm10 C-terminus, amino acid D82 of FgPre6 and K62 of FgPre5. Additional FgBlm10-interacting proteins were identified in the wild type and pkr mutant, suggesting that PKA regulates the preference of FgBlm10-mediated proteasome assembly. In addition, PKA indirectly affected the phosphorylation of FgBlm10, and its localization in the nucleus. The truncation of the FgBlm10 C terminus also enhanced nuclear import and bleomycin resistance, suggesting its role in proteasome assembly at DNA damage sites. Collectively, our data demonstrated that regulation between PKA and proteasome degradation is critical for the vegetative growth of F. graminearum

Comparative Study on Connection Properties of Shear Bolt and Screw of Thin Cross-laminated Timber Panel

Cross-laminated timber (CLT), a wood product with excellent shear resistance, is often used in modern timber constructions. Using the standards ASTM D1761-12 (2020) and NDS-2012 (2012), this study investigated the connection properties of shear bolts and screws in CLT panels. The specimens were made from spruce-pine-fir lumber and installed on a test platform using one high-strength bolt or eight screws, and then an upward load was applied to the top of the specimen. The results showed that the bolt connection provided a higher ultimate bearing capacity and elastic stiffness. The bolt exhibited virtually no deformation, and the CLT panel did not noticeably deteriorate when the connection was damaged. The distance between the bolt hole and the bottom of the CLT specimen and the angle between the outer-layer grain direction of the CLT panel and the load direction were both measured. Changes in the ductility coefficient value had an obvious effect on the connection performance of the shear bolts when the outer-layer grain direction of the CLT panel was consistent with the load direction. Contrastingly, when the outer-layer grain direction of the CLT panel was perpendicular to the load direction, the effect was negligible, and the yield load was nearly unchanged