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        Molecular cloning, characterization, and functional analysis of a gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A synthase from Matricaria chamomilla

        Feng Xu,Tingting Tao,Qiangwen Chen,Xiangxiang Meng,Jiaping Yan,Jie Chang 한국유전학회 2016 Genes & Genomics Vol.38 No.12

        3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) catalyzes the condensation of acetyl-CoA and acetoacetyl- CoA to form 3-hydroxy-3-methylglutaryl-CoA as the first committed enzyme in the mevalonate (MVA) pathway. HMGS plays an important role in the biosynthesis of the sesquiterpene, which is the main constituent of essential oil in Matricaria chamomilla. In this paper, a HMGS gene designated as McHMGS (GenBank Accession No. KU529970) was successfully cloned from M. chamomilla. The full-length cDNA of McHMGS was 1495-bp and contained a 1374-bp open reading frame. It encoded a 458-amino-acid protein with a calculated molecular weight of about 50.7 kDa and isoelectric point of 5.69. Sequence comparison revealed that McHMGS showed extensive homology with HMGSs from other plant species. Phylogenetic tree analysis indicated that McHMGS is clustered with the HMGS of Asteraceae in the dicotyledoneae clade. Further functional complementation of McHMGS in hmgsdeficient mutant yeast strain YSC6274 demonstrated that cloned McHMGS cDNA encodes a functional HMGS and mediates the MVA biosynthesis in yeasts. The tissue expression pattern analysis revealed that McHMGS expression level is highest in the flowers and lowest in the stems. Quantitative real-time PCR analysis showed that the expression of McHMGS was induced by MeJA, and the expression level is highest 24 h after induction. The characterization and expression of McHMGS can help in further studying the role of McHMGS gene in the biosynthesis of sesquiterpene in M. chamomilla.

      • Analysis of shear lag effect in the negative moment region of steel-concrete composite beams under fatigue load

        Jinquan Zhang,Bing Han,Hui-bing Xie,Wu Tong Yan,Wangwang Li,Jiaping Yu 국제구조공학회 2021 Steel and Composite Structures, An International J Vol.39 No.4

        Shear lag effect was a significant mechanical behavior of steel-concrete composite beams, and the effective flange width was needed to consider this effect. However, the effective flange width is mostly determined by static load test. The cyclic vehicle loading cases, which is more practical, was not well considered. This paper focuses on the study of shear lag effect of the concrete slab in the negative moment region under fatigue cyclic load. Two specimens of two-span steel-concrete composite beams were tested under fatigue load and static load respectively to compare the differences in the negative moment region. The reinforcement strain in the negative moment region was measured and the stress was also analyzed under different loads. Based on the OpenSees framework, finite element analysis model of steel-concrete composite beam is established, which is used to simulate transverse reinforcement stress distribution as well as the variation trends under fatigue cycles. With the established model, effects of fatigue stress amplitude, flange width to span ratio, concrete slab thickness and shear connector stiffness on the shear lag effect of concrete slab in negative moment area are analyzed, and the effective flange width ratio of concrete slab under different working conditions is calculated. The simulated results of effective flange width are compared with calculated results of the commonly used specifications, and it is found that the methods in the specifications can better estimate the shear lag effect in concrete slab under static load, but the effective flange width in the negative moment zone under fatigue load has a large deviation.

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