Targeting of human aFGF gene into silkworm, Bombyx mori L.Through homologous recombination

The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination.     

Targeting of human aFGF gene into silkworm,<Emphasis Type="Italic">Bombyx mori</Emphasis> L. through homologous recombination

The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm,Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL 1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination. Project supported by the Scientific Research Foundation for Returned Overseas Chinese Scholars, Education Ministry of China and the Natural Science Foundation of Zhejiang Province (No. 301306), China     

昆虫杆状病毒在生物技术研究中的应用

随着昆虫杆状病毒分子生物学研究的不断深入，昆虫杆状病毒在生物技术研究中也得到了应用。利用杆状病毒作为载体，在昆虫细胞和虫体内表达外源基因，形成了昆虫杆状病毒载体表达系统，利用杆状病毒携带外源基因，通过同源重组将外源基因导入到家蚕的基因组构建了转基因家蚕；通过向杆状病毒基因组中插入毒素、激素、酶抑制剂等的基因或从杆状病毒基因组中删除某个基因，或通过不同杆状病毒间的杂交，使子代病毒的宿主域扩大，从而提高了杆状病毒作为生物杀虫剂的杀虫效果。本文对杆状病毒在上述几个方面的应用进行了简要综述。     

秦巴山区桑蚕野生资源的开发与利用

野桑蚕是与家蚕亲缘的野生资源,是家蚕遗传进化研究的物质基础,是家蚕品种选育重要的种质资源.秦巴山区是野桑蚕的起源地,独特的地理位置和气候条件,进化形成了丰富的野桑蚕种质类别.初步查明秦巴山区野桑蚕的多态性表现极为丰富,其中有着重大开发利用前景的有三个方向:一是野桑蚕天然有色茧基因的分离和转育;二是野桑蚕抗性基因利用;三是野桑蚕特异性生态变异的研究.     

昆虫的生物测时机构

生物测时是昆虫的基本生理功能。昆虫测时功能及作用机制的研究是时间生物学前沿研究内容之一。光感应型昆虫生物钟光信号由眼、脑－眼两种感知类型,驱动部位也有视叶边缘腹侧部分的细胞体和脑两类;遗传控制基因是ｐｅｒ,不同突变体ｐｅｒＤＮＡ之间仅有１个碱基的差异。家蚕卵内的酯酶Ａ４（ＥＡ４）是一种时间间隔测定酶,感知低温、盐酸刺激等信息,启动倒计时型滞育生物钟光ＥＡ４的测时功能与自身的结构有关,受蚕卵内的多肽     

麻风树核糖体失活蛋白的基因组DNA分离及其转化烟草分析

1IntroductionRibosome-inactivating proteins(RIPs)occur natu-rally in a variety of higher plant species,and theyfunction by catalytic depurination of a specific aden-osine residue located near the3*terminus of eukary-otic large ribosomal subunit rRNA,preventing EF-2/GTP binding and thereby blocking peptidyl-tRNAtranslocation during protein synthesis[1].Many RIPsare potent antiviral and antifungal proteins in vitro[2,3],but it may beinsufficient for field application,as com-pared to the ac…     

苏南老蚕区丝绸产业链转型升级策略

在当今国际贸易形势和国家"东桑西移"的政策背景下,传统茧丝绸产业面临着新的挑战。明确茧丝绸业"苏南模式"的新内涵和存在的必要性,正视老蚕区在产业结构调整过程中暴露的劳动力不足、茧丝原材料质量下降等诸多"短板",深入探讨苏南老蚕区茧丝绸产业链转型升级发展的相关对策,具有十分重要的历史意义。     

简并PCR法克隆L.starkeyi苹果酸酶基因

根据已报道的酵母苹果酸酶基因序列,利用CodeHop（Consensus Degenerate Hybrid Oligonucleotide Primers）软件设计两对简并引物,以斯达油脂酵母（L.starkeyi CICC 1809）总DNA为模板做简并PCR,得到2个基因片段（ME1,ME2）.对这2个目的片段进行测序,将结果翻译成氨基酸序列,blastp结果显示其中ME2片段为油脂酵母未报道苹果酸酶基因的序列（Gene Bank登录号GU348991）,并对巢式PCR方法检验简并PCR结果的有效性进行了评估.     

Using chimeric piggyBac transposase to achieve directed interplasmid transposition in silkworm Bombyx mori and fruit fly Drosophila cells

The piggyBac transposon has been long used to integrate foreign DNA into insect genomes. However, undesirable transgene expression can result from random insertions into the genome. In this study, the efficiency of chimeric Gal4-piggyBac transposase in directing integration onto a DNA target plasmid was evaluated in cultured silkworm Bombyx mori Bm-12 and fruit fly Drosophila Schneider 2 (S2) cells. The Gal4-piggyBac transposase has a Gal4 DNA-binding domain (DBD), and the target plasmid has upstream activating sequences (UAS) to which the Gal4 DBD can bind with high affinity. The results indicate that, in the Bm-12 and S2 cells, transpositional activity of Gal4-piggyBac transposase was increased by 4.0 and 7.5 times, respectively, compared to controls, where Gal4-UAS interaction was absent. Moreover, the Gal4-piggyBac transposase had the ability of directing piggyBac element integration to certain sites of the target plasmid, although the target-directing specificity was not as high as expected. The chimeric piggyBac transposase has the potential for use in site-directed transgenesis and gene function research in B. mori.     

DNA芯片技术及其应用

DNA芯片技术是近年发展起来的DNA分析技术，它采用高速打印或光刻合成技术可在硅片、玻璃或尼龙膜上制造DNA微阵列.样品DNA通过PCR扩增、体外转录等技术掺入荧光标记分子，与微阵列杂交后，通过荧光扫描仪扫描及计算机分析即可获得样品中大量基因序列及表达信息.该技术可应用于DNA序列测定、基因表达水平检测、基因及多态性检测、发现新基因等多个研究领域.     

论家蚕传染病的综合防治

家蚕传染病的发生和蔓延,是由病原体侵染、蚕体的生理状况和环境条件等多方面因素共同作用的结果。本文根据各类蚕病发生规律,对家蚕传染性蚕病的综合防治措施进行了全面探讨。     

清末蚕桑教育的兴办及其影响

蚕桑教育在清末备受重视,蚕桑学堂、蚕桑讲习所、蚕桑传习所以及设有蚕业专科的农业学堂纷纷设立,清末的蚕桑教育取得了令人瞩目的进展.清末蚕桑教育的兴办,为各地培养了大批懂得近代蚕桑技术的专门人才,促进了各地蚕桑新知识技术的推广和传播,推动了蚕桑业的改良和发展.     

EFFECTS OF JINLU, AN ANTI-JUVENILE HORMONE ON THE GROWTH, ULTRA-STRUCTURE OF THE CORPORA ALLATA AND PROTHORACIC GLAND OF SILKWORM, BOMBYX MORIL

ＩＮＴＲＯＤＵＣＴＩＯＮＩｎｓｅｃｔｐｏｓｔｅｍｂｒｙｏｎｉｃｄｅｖｅｌｏｐｍｅｎｔｃｏｎｓｉｓｔｓｏｆｇｒｏｗｔｈｐｕｎｃｔｕａｔｅｄｂｙａｓｅｒｉｅｓｏｆｍｏｌｔｓｆｏｌｌｏｗｅｄｂｙｍｅｔａｍｏｒｐｈｏｓｉｓ.Ｔｈｅｓｅｍｏｌｔｓａｎｄｍｅｔａｍｏｒ ｐｈｏｓｉｓａｒｅｉｎｉｔｉａｔｅｄａｎｄｃｏｏｒｄｉｎａｔｅｄｂｙｈｏｒｍｏｎｅｓ(Ｒｉｄｄｉｆｏｒｄ ,1 994 ) .Ｉｔｉｓｇｅｎｅｒａｌｌｙａｃｃｅｐｔｅｄｔｈａｔｔｈｅｉｎｔｅｒｐｌａｙｏｆｅｃｄｙｓｔｅｒｉｏｄｓ,ａｇｒｏｕｐｏｆｓｔｅｒｏ…     

乙酰短杆菌α-乙酰乳酸脱羧酶基因的克隆及其表达

用PCR方法扩增乙酰短杆菌α-乙酰乳酸脱羧酶基因(ALDC),得到0.78 kb的DNA片段,扩增片段重组到表达载体pQE-9中,在大肠杆菌中得到高效表达,酶活性可达100 U/mL发酵液.     

Computational prediction of microRNA genes in silkworm genome

MicroRNAs （miRNAs） constitute a novel, extensive class of small RNAs （-21 nucleotides）, and play important gene-regulation roles during growth and development in various organisms. Here we conducted a homology search to identify homologs of previously validated miRNAs from silkworm genome. We identified 24 potential miRNA genes, and gave each of them a name according to the common criteria. Interestingly, we found that a great number of newly identified miRNAs were conserved in silkworm and Drosophila, and family alignment revealed that miRNA families might possess single nucleotide polymorphisms, miRNA gene clusters and possible functions of complement miRNA pairs are discussed.     

Accumulation of 1-deoxynojirimycin in silkworm, <Emphasis Type="Italic">Bombyx mori</Emphasis> L.

1-deoxynojirimycin (1-DNJ) contents in the silkworm,Bombyx mori,at different developmental stages and tissues were investigated by using reverse-phase high-performance liquid chromatography. The 1-DNJ contents of silkworm larvae change significantly with their developmental stages. The male larvae showed higher accumulation efficiency of 1-DNJ than the females and also a significant variation was observed among the silkworm strains. The present results show that tissue distribution of 1-DNJ was significantl...     

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtilis was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone α-factor (MFals), and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction (PCR)-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae (SC-βG) was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A (PrA) activity was measured in fermenting liquor. The results of PCR analysis ofgenome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/(h-ml) after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.     

辽宁柞蚕纺织产业集群发展的分化与协作

制度性因素可以导致集群发展的分化,辽宁丹东与盖州的柞蚕纺织产业集群发展印证了这一点。针对辽宁柞蚕纺织产业集群分布的地区优势,建议整合资源,给予相应政策扶持;延伸柞蚕纺织产业链条;借力桑蚕丝织品销售网络和渠道;展开国际间技术合作,以便节约集群企业交易成本,促进集群的可持续发展。     

萤火虫萤光素酶基因转化花叶芋及表达的研究

将萤火虫萤光素酶基因重组到质粒pBR325中,通过“三亲交配”和同源重组,将萤光素酶基因整合到Ti质粒pGV3850:1103。通过改良的叶盘转化法,用萤光素酶基因转化单子叶植物花叶芋,获得转基因植株。对再生植株的胭脂碱测定、新霉素磷酸转移酶分析、DNA点杂交、萤光素酶基因活性测定,证明萤光素酶基因已转化到花叶芋中并进行了表达。