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| 1 | Construction of a host range-expanded hybrid baculovirus of BmNPV and AcNPV, and knockout of cysteinase gene for more efficient expression显示文摘AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV(Bombyx mori nuclear polyhedrosis virus) are two principal insect-baculovirus expression systems, each having different characteristics. AcNPV has a wider host range and can infect a series of cell lines thus making it suitable for cell suspension culture expression, but the small size of the host insect, A. californica, makes AcNPV less suitable for large scale protein synthesis. In contrast, BmNPV can only infect the silkworm, Bombyx mori, which is well-known for its easy rearing and large size. These characteristics make the BmNPV system especially suitable for large-scale industrial ex-pression. To utilize the advantages of both AcNPV and BmNPV, we tried to expand their host range through homologous recombination and successfully constructed a hybrid baculovirus of AcNPV and BmNPV, designated as HyNPV. The hybrid baculovirus can infect the hosts of both AcNPV and BmNPV. Taking the human basic fibroblast growth factor (bFGF) gene as an appli-cation example, we constructed a recombinant, HyNPV-bFGF. This construct is able to express the bFGF protein both in silkworm larvae and in common-use cell lines, sf21, sf9 and High-five. Moreover, to reduce the loss of recombinant protein due to degradation by proteases that are simultaneously expressed by the baculovirus, we knocked out the cysteinase gene coding for one of the most important baculovirus proteases. This knockout mutation improves the produc-tion efficiency of the bFGF recombinant protein. | WU Xiaofeng, CAO Cuiping, XU Yaxiang & LU Xingmeng College of Animal Sciences, Zhejiang University, Huajiachi Campus, Hangzhou 310029, China | 2004 | Science China(Life Sciences)2004，47，5： | 12 |
| 2 | Transcriptome sequencing and characterization of ungerminated and germinated spores of Nosema bombycis显示文摘Nosema bombycis 是一 obligate 利用特殊机制感染 Bombyx mori 的细胞内部的寄生真菌。萌芽,通过哪个 microsporidia 感染主人房间的一个 indispensible 过程,从休眠状态为 microsporidia 被认为是一个关键发展拐弯处点到繁殖状态。因此,阐明在萌芽为寄生虫控制是关键的前后, transcriptome 变化。然而, microsporidia 的萌芽的分子的基础仍然保持未知。调查这个萌芽过程, N 的 transcriptome。bombycis ungerminated 孢子和发芽孢子被定序并且分析。超过 6000 万个高质量的抄本读用 RNA-Seq 技术从这二个组被产生。在汇编以后,分别地, 2756 和 2690 unigenes 被识别并且随后基于已知的蛋白质注解了。在差别的分析表示了基因以后, 66 基因被识别是在这二个组之间的表示的差别(P 0.05 ) 。蛋白质联系磷酸酶的基因是由 RNA-Seq 和 immunoblot 分析决定了首先被识别显著地起来调整,显示那 dephosphorylation 可能潜在地贡献 microsporidia 萌芽。编码涉及 glycometabolism,孢子墙蛋白质和 N 的篦麻毒素 B lectin 的蛋白质的 DEG。bombycis 也被分析。基因和染色体分析的基因本体论和京都百科全书揭示了为一些特定的生物函数和进程负责的基因。在这研究产生的数据集在 N 提供 transcriptome 变化的基本描述。bombycis 在萌芽期间。transcriptome 数据的分析和是与萌芽有关的柔韧的候选人基因的某些功能的基因的鉴定将帮助提供孢子萌芽和侵略的深理解。 | Han Liu Mingqian Li Xinyi He Shunfeng Cai Xiangkang He Xingmeng Lu | 2016 | Acta Biochimica et Biophysica Sinica2016，48，3： | 10 |
| 3 | Analysis of ESTs and gene expression patterns of the posterior silkgland in the fifth instar larvae of silkworm, Bombyx mori L.显示文摘The fibroin gene expression pattern and regulation of the posterior silkgland were studied by means of expressed sequence tags (ESTs) using the first and fifth day larvae of the fifth instar of silkworm, Bombyx mori L (strain: C 108). The results showed that there were 911 repetitive ESTs and 1950 single sequences (Singlets) among total 2861 consentient sequences, which were spliced. 1335 sequences were identified and the other 1526 were unknown. 5560 sequences (55.89%) in the posterior silkgland cell of the silkworm were new ESTs without homology with EST data published by Mita et al. The number of repetitive ESTs and single sequences from the first day larvae of the fifth instar was double more than that of the fifth day of the same instar in the silkworms. The unigenes which were more than 50 in repetitive EST size (contig size) came to only about 0.5% in total consentient sequences. There were significant differences between gene expression frequencies, and expressed genes were related to fibroin synthesis and its secretion and fibroin composition. Comparing the fifth day with the first day of the fifth instar, the genes-expressed quantity of fibroin heavy-chain gene was 18 fold higher, fibroin light-chain gene 9 fold and fibroin P52 gene 8 fold. 508 genes functioned for cellular component and 315 for enzyme after function tracing. These results implied that the gene expression of the first day was mainly for preparation for fibroin synthesis except for the growth of silkgland cells, and the gene expression of the fifth day of the fifth instar was mainly for synthesizing and excreting fibroin. Because the ratio of heavy chain, light chain and p25 of fibroin was not 6:6:1 as theoretically expected, or its special H-chain structure, the H-chain gene was not easy to detect through EST technique. Most of genes among total 2861 consentient sequences functioned for fibroin synthesis and secretion. This suggested the fibroin synthesis and secretion procedure of the posterior silkgland was more complex than the knowledge we have. | ZHONG Boxiong, YU Yingpo, XU Yusong, YU Hong, LU Xingmeng, MIAO Yungen, YANG Jun, XU Hao, HU Songnian & LOU Chengfu College of Animal Sciences, Zhejiang University, Hangzhou 310029, China Hangzhou Huada Gene Research and Development Center, Southern Base of Genomic Information, Chinese Academy of Sciences, Hangzhou 310007, China | 2005 | Science China(Life Sciences)2005，48，1： | 8 |
| 4 | Ricin-B-lectin enhances microsporidia Nosema bombycis infection in BmN cells from silkworm Bombyx mori显示文摘 | Han Liu Mingqian Li Shunfeng Cai Xinyi He Yongqi Shao Xingmeng Lu | 2016 | Acta Biochimica et Biophysica Sinica2016，48，11： | 6 |
| 5 | Proteomic analysis of BmN cells (Bombyx morl) in response to infection with Nosema bombycis显示文摘Nosema bombycis (N。bombycis, Nb ) 一 obligate 细胞内部的寄生虫,它能在蚕引起蚕胞子虫病疾病。调查 N 的效果。主人房间上的 bombycis 感染,从没感染 N 的 BmN 房间的 proteomes。在不同感染阶段的 bombycis 与二维的胶化电气泳动和 MALDI-TOF/TOF 集体 spectrometry 被描绘,它鉴别 24 差别与重要紧张差别表示了主人蛋白质(P < 0.05 ) 同时至少指在嘲笑 -- 并且 N。bombycis 感染了房间。尤其是,基因本体论分析证明这些蛋白质涉及许多重要生物反应。在感染阶段期间,涉及精力新陈代谢和氧化应力的蛋白质有起来调整的表示。二蛋白质参予了 ubiquitin 依赖的蛋白质分解代谢的过程有的下面调整的表示。量的即时聚合酶链反应被用来分析这些识别蛋白质的 transcriptional 侧面。一起拿,在为识别蛋白质的生物反应的丰富变化,通常认为的功能,和参予在 N 生产一个主人应答的蛋白质模型。感染 bombycis 的 BmN 房间。这些调查结果在宿主细胞上关于精力缺点寄生虫的效果推进我们的知识。 | Xinyi He Xiangkang He Han Liu Mingqian Li Shunfeng Cai Zhangwuke Fu Xingmeng Lu | 2014 | Acta Biochimica et Biophysica Sinica2014，46，11： | 5 |
| 6 | Nosema bombycis (Microsporidia) suppresses apoptosis in BmN cells (Bombyx mori)显示文摘Nosema bombycis (N。bombycis, Nb ) 一真菌相关并且要求在蚕引起长期的蚕胞子虫病疾病的细胞内部的寄生虫。在感染主人以后,孢子从主人房间获得精力并且幸存几天。在病原体和宿主细胞之间的这共生建议那 N。bombycis 阻止 apoptosis 和宿主细胞的反应的氧种(ROS ) 生产为它的生长和发展创造最佳的环境条件。在这研究,不同方法被用来证明那 N。bombycis 在 BmN 房间压制了 apoptosis。流动 cytometry 分析结果证明孢子在感染以后在 2 和 5 天压制了 BmN 房间的 apoptosis (P < 0.05 ) 。与放线菌素 D (ActD ) 处理相比, BmN 房间的 apoptosis 显然在孢子感染以后被减少(P < 0.01 ) 。在感染以后的 48 个小时, BmN 房间的 ROS 生产在为 6 h 的 ActD 处理以后与那相比是下面调整的。而且, N。bombycis 阻止了 apoptosomes 的形成由下面调整 apaf-1 和细胞色素 C 的表示。另外, N。bombycis 也起来调整 buffy 的表示。西方的污点分析证明孢子减少了在 48 和 98 h 柱子感染的主人细胞色素 C 的水平。因此,我们的结果建议了那 N。bombycis 禁止了主人房间的 mitochondrial apoptotic 小径为它的自己的幸存创造最佳的环境。 | Xinyi He Zhangwuke Fu Mingqian Li Han Liu Shunfeng Cai Nana Man Xingmeng Lu | 2015 | Acta Biochimica et Biophysica Sinica2015，47，9： | 4 |
| 7 | Gene analysis of an antiviral protein SP-2 from Chinese wild silkworm, Bombyx mandarina Moore and its bioactivity assay显示文摘The cDNA encoding an antiviral protein SP-2 against BmNPV was cloned from the midgut of Chinese wild silkworm, Bombyx mandarina Moore (GenBank access AY945210) based on the available informa- tion of the domesticated silkworm. Its cDNA was 855 bp encoding 284 amino acids with predicted mo- lecular weight of 29.6 kDa. Its full length in genomics was 1376 bp, including 5 exons and 4 introns. The expression analysis indicated that it was only expressed in midgut, and its expression level was higher during feeding stage of larval instars while very lower during the moltism and mature stages. The de- duced amino acid sequence of this protein showed eight-amino-acid variation compared with the counterpart of domesticated silkworm. Its antiviral activity was assayed through in vitro test. The re- sults indicated that it showed strong bioactivity against BmNPV, and its activity was 1.6 fold higher that the counterpart of domesticated silkworm. | YAO HuiPeng, HE FangQing, GUO AiQin, CAO CuiPing, LU XingMeng & WU XiaoFeng College of Animal Science, Zhejiang University, Hangzhou 310029, China | 2008 | Science China(Life Sciences)2008，51，10： | 2 |
| 8 | Taxonomical studies on enterococci isolated from the intestine of the silkworm,Bombyx mori显示文摘 | Lu Xingmeng Hashimoto Y Shimizu S | 1994 | J of Sericultural Science of Japan1994，63，： | 1 |
| 9 | Proteomic analysis of silkworm midgut cellular proteins interacting with the 5' end of infectious flacherie virus genomic RNA显示文摘在蚕的 flacherie 疾病被传染 flacherie 病毒(IFV ) 引起。IFV 依靠它 genomic RNA 的 5 区域到成员实现病毒的翻译和复制的主人相关的因素。为了识别,招待 IFV RNA 的 5 区域一定的蛋白质并且识别为它的函数重要的蛋白质,集体 spectrometry 被用来从用 RNA 被获得的蚕 midgut 摘录识别蛋白质标记 aptamer IFV RNA 的 5 区域。我们发现 325 蛋白质组(唯一的肽 2 )包括翻译相关的因素的 IFV RNA 的 5 区域一定( 16 个 ribosomal 子单元, 3 个真核细胞的开始因素子单元, 1 个延伸因素子单元和 6 潜力内部核糖体入口地点交易因素),细胞骨架相关的蛋白质,膜相关的蛋白质,新陈代谢酶,并且另外的蛋白质。这些结果能被用来学习翻译,复制联系了在蚕与主人蚕并且到控制 flacherie 疾病交往的 IFV 的因素。 | Mingqian Li Xinvi He Han Liu Zhangwuke Fu Xiangkang He Xingmeng Lu | 2015 | Acta Biochimica et Biophysica Sinica2015，47，2： | 0 |
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