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Knockout of the EgriBLOS2 gene results in the transparent integuments of Ectropis grisescens larvae
Li Jia-Li,Zhuang Xiang-Lin,Yuan Ting-Ting,Cai Xiao-Ming,Luo Zong-Xiu,Bian Lei,Chen Zong-Mao,Li Zhao-Qun,Liu Nai-Yong 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.1
The body colors of insects have evolved specialized roles in body protection, signaling and physiology. In some case, the larvae can camouflage their body colors to adapt the habitants and protect themselves. However, the genetic and molecular basis on larval body colors of the tea geometrid, Ectropis grisescens, remains poorly known. Here, we reported an effect of the lysosome-related organelles complex-1, subunit 2 (EgriBLOS2) gene knockout on larval integuments of E. grisescens, by using a clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system. Combining transcriptomic data and PCR approaches, we first identified the EgriBLOS2 gene from E. grisescens, which encoded 137 amino acids and comprised 3 introns. In the phylogenetic analysis, EgriBLOS2 clustered in the Lepidoptera clade with high conservation to members in other lepidopteran species. Developmental expression profiles revealed that EgriBLOS2 was constantly tran scribed at each stage, in which its expression was significantly lower in 2nd instar larvae than that of other instars. After injecting EgriBLOS2-specific guide RNA and Cas9 messenger RNA into eggs, 70% of larvae showed the translucent integuments in G0 generation, with an emphasis on black splayed patterns in the 2nd and 8th segments of abdomens. However, some typical characteristics of larvae were not obviously changed in mutant instars, such as ocelli, mouthparts and other appendants. This study has unraveled the roles of EgriBLOS2 in the formation of larval integument colors, and provides an alternative strategy for pest management based on the colors in this species.
Xiang Zou,Chang-fa Chen,Xia-chang Qi,Jiang-chao Qian,Ju Chu,Ying-ping Zhuang,Si-liang Zhang,Wei Zeng,Wan-jun Li 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.6
Improvement of Erythromycin A (Er-A) production and purity by metabolic engineering of the industrial erythromycin-producing strains Saccharopolyspora erythraea strians ZL1004 and ZL1007, in which the amounts of tailoring enzymes EryK (a P450 hydroxylase)and EryG (an S-adenosylmethionine-dependent O-methyltransferase)for biotransformation of Erythromycin D to Er-A were modulated, was performed in a 50 L fermentor. Addition of 15 g/L of corn steep liquor to the medium increased Er-A production; maximum Er-A production was 8,196 U/mL at 191 h, which was 81.8% higher than that of control (4,507 U/mL at 184 h). Er-B impurities were completely eliminated, whereas Er-C impurities were only 153 U/mL at 191 h. Analysis of intra- and extracellular metabolites and key enzyme activities in central carbon metabolism revealed that the pool of TCA cycle intermediates was enhanced by the addition of corn steep liquor and induced an increase in erythromycin biosynthesis. There were no significant differences between strains ZL1004 and ZL1007 regarding Er-A production and impurity accumulation. Compared to wild type strain,Er-A production was improved by 23.9% while Er-C was reduced by 83.9% and Er-B was completely eliminated. Furthermore, fermentation of recombinant strain ZL1004was successfully scaled up from laboratory scale (50 L fermentor) to industrial scale (25 and 132 m3), with similar levels of Er-A production and purity obtained.
Minnan Yang,Zhuang Ding,Qun Xiang,Xiaodong Zhang,Xiang Li,Seydou Sylla 한국미생물학회 2014 The journal of microbiology Vol.52 No.4
Porcine reproductive and respiratory syndrome (PRRS) is animportant disease, which leads to severe economic losses inswine-producing areas of the world. However, current antiviralstrategies cannot provide highly effective protection. In this study, three theoretically effective interference targetsites (71-91, 144-164, 218-238) targeting the nucleocapsid (N)gene of PRRSV were designed and selected, and then threesiRNA-expressing plasmids were constructed, respectivelynamed p2.1-N71, p2.1-N144, and p2.1-N218. The recombinantsiRNA-expressing plasmids were transfected into Marc-145 cells; then the cells were infected with PRRSV (JL07SWstrain); finally, after incubation for 48 h, the antiviral activityof those siRNA-expressing plasmids in Marc-145 cells wasassessed by cytopathic effects, virus titers, indirect immunofluorescence,and quantitative real-time PCR. Experimentalresults demonstrated that these three siRNA-expressing plasmidscould effectively and significantly inhibit the replicationof PRRSV by 93.2%, 83.6%, and 89.2% in Marc-145 cells,respectively. Among these three siRNA-expressing plasmids,p2.1-N71 was found to be most effective, while p2.1-N144and p2.1-N218 displayed relatively weak inhibition of virusreplication. The results indicated that siRNA-expressing plasmidstargeting the N gene of PRRSV could significantly inhibitPRRSV replication in Marc-145 cells. Based on our experimentalresults and previous reports, the 71-91, 179-197,and 234-252 sites of the N gene are good choices to effectivelyinhibit the replication of PRRSV, and this RNA interferencetechnique can be a potential anti-PRRSV strategy.
Static and fatigue behavior of through-bolt shear connectors with prefabricated HFRC slabs
Yuliang He,Jie Zhuang,Lipu Hu,Fuyou Li,Ying Yang,Yi-qiang Xiang 국제구조공학회 2022 Structural Engineering and Mechanics, An Int'l Jou Vol.83 No.1
Twelve push-out test specimens were conducted with various parameters to study the static and fatigue performance of a new through-bolt shear connector transferring the shear forces of interface between prefabricated hybrid fiber reinforced concrete (HFRC) slabs and steel girders. It was found that the fibers could improve the fatigue life, capacity and initial stiffness of through-bolt shear connector. While the bolt-hole clearance reduced, the initial stiffness, capacity and slippage of through-bolt shear connector increased. After the steel-concrete interface properties were improved, the initial stiffness increased, and the capacity and slippage reduced. Base on the test results, the equation of the load-slip curve and capacity of through-bolt shear connector with prefabricated HFRC slab were obtained by the regression of test results, and the allowable range of shear force under fatigue load was recommended, which could provide the reference in the design of through-bolt shear connector with prefabricated HFRC slabs.
Wu Jiang,Fu Liwei,Yan Zineng,Yang Yu,Yin Han,Li Pinxue,Yuan Xun,Ding Zhengang,Kang Teng,Tian Zhuang,Liao Zhiyao,Tian Guangzhao,Ning Chao,Li Yuguo,Sui Xiang,Chen Mingxue,Liu Shuyun,Guo Quanyi 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00
In recent years, there has been significant research progress on in situ articular cartilage (AC) tissue engineering with endogenous stem cells, which uses biological materials or bioactive factors to improve the regeneration microenvironment and recruit more endogenous stem cells from the joint cavity to the defect area to promote cartilage regeneration.In this study, we used ECM alone as a bioink in low-temperature deposition manufacturing (LDM) 3D printing and then successfully fabricated a hierarchical porous ECM scaffold incorporating GDF-5.Comparative in vitro experiments showed that the 7% ECM scaffolds had the best biocompatibility. After the addition of GDF-5 protein, the ECM scaffolds significantly improved bone marrow mesenchymal stem cell (BMSC) migration and chondrogenic differentiation. Most importantly, the in vivo results showed that the ECM/GDF-5 scaffold significantly enhanced in situ cartilage repair.In conclusion, this study reports the construction of a new scaffold based on the concept of in situ regeneration, and we believe that our findings will provide a new treatment strategy for AC defect repair.