麦红吸浆虫滞育发生和解除过程中总脂和甘油三酯含量变化

【目的】脂类与昆虫滞育密切相关。本研究旨在明确麦红吸浆虫Sitodiplosis mosellana (Géhin)滞育过程中脂类物质含量的变化规律,探讨麦红吸浆虫滞育与脂类物质变化的关系。【方法】采用香兰素硫酸显色法测定了2008年5月－2010年1月不同时间陕西杨凌养虫圃内麦红吸浆虫滞育前、滞育期及滞育解除后幼虫和蛹体内总脂和甘油三酯含量。【结果】 滞育前幼虫总脂和甘油三酯含量分别为378.12和291.67 μg/mg,显著高于整个滞育期（P<0.05）。滞育年周期中,冬季总脂和甘油三酯含量最高,翌年1月结茧幼虫二者含量分别为335.29 和275.72 μg/mg,显著高于其他季节（P<0.05）;整个滞育期间,裸露幼虫和结茧幼虫总脂和甘油三酯随季节变化趋势相同,但同期结茧幼虫含量高于裸露幼虫;滞育当年与第2年同期幼虫总脂和甘油三酯含量差异不显著（P>0.05）。滞育解除后,总脂和甘油三酯含量随着幼虫的发育和变态逐渐降低,其中中蛹和后蛹显著低于活动幼虫（P<0.05）。【结论】麦红吸浆虫滞育不同时期幼虫及蛹总脂和甘油三酯含量存在明显差异,其滞育与脂类物质的含量密切相关。     

低温对麦红吸浆虫滞育解除及蜕皮激素受体基因EcR和热激蛋白基因Hsp70和Hsp90表达水平的影响

【目的】本研究旨在明确破茧率和破茧所需时间作为典型的专性幼虫期滞育昆虫麦红吸浆虫Sitodiplosis mosellana滞育解除指标的可行性,探讨蜕皮激素受体基因EcR和热激蛋白基因Hsp70和Hsp90在低温解除滞育中的作用。【方法】9月上旬采自田间的麦红吸浆虫滞育幼虫在低温(4℃)和自然变温处理不同时间(0~90 d)后转移至24℃化蛹,调查幼虫的破茧率、化蛹率及破茧和化蛹所需时间;采用qPCR技术分析4℃低温终止滞育并在24℃下恢复发育的幼虫EcR, Hsp70和Hsp90的mRNA水平。【结果】不同时间低温(4℃)和自然变温处理的麦红吸浆虫滞育幼虫破茧率和化蛹率以及破茧和化蛹所需时间均存在显著差异。未经低温(4℃)处理的幼虫在水分满足的条件下破茧率为71.3%,但均不能化蛹;低温(4℃)处理60 d内,随着处理时间的延长破茧率和化蛹率逐渐升高,破茧和化蛹所需时间逐渐缩短;与4℃低温处理30~60 d相比,自然变温处理30~60 d的幼虫化蛹率显著较低;4℃低温处理60 d和自然变温处理90 d后幼虫的化蛹率均超过91%。低温(4℃)处理显著提高了麦红吸浆虫幼虫EcR, Hsp70和Hsp90的表达量,其中处理30 d时其表达量最高,随着低温(4℃)处理时间的延长,表达量逐渐降低,大部分幼虫滞育解除后表达量趋于恒定。【结论】低温能显著促进麦红吸浆虫的滞育解除,效果优于9-10月自然变温;破茧率及破茧所需时间可作为评判麦红吸浆虫幼虫滞育强度的参考,但不能独立作为滞育解除的指标;EcR,Hsp70和Hsp90的表达水平与滞育强度密切相关,在麦红吸浆虫滞育解除中发挥潜在作用。     

麦红吸浆虫3-羟基-3-甲基戊二酰辅酶A还原酶基因SmHMGR的克隆及在滞育与发育变态过程中的表达动态

【目的】3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)是保幼激素(JH)合成途径的限速酶。麦红吸浆虫Sitodiplosis mosellana是一种典型的专性幼虫滞育昆虫。本研究旨在探讨HMGR基因在麦红吸浆虫滞育和发育变态过程中的作用。【方法】通过RT-PCR和RACE技术克隆麦红吸浆虫滞育前幼虫HMGR基因全长cDNA序列;利用生物信息学软件分析HMGR基因核苷酸和其编码的蛋白氨基酸序列特性;采用qPCR技术测定其在麦红吸浆虫滞育不同时期3龄幼虫及不同发育阶段(1-2龄幼虫、预蛹、初蛹、中蛹和后蛹以及雌雄成虫)中的mRNA表达水平。【结果】克隆获得一条麦红吸浆虫HMGR基因全长cDNA序列,命名为SmHMGR(GenBank登录号: MG876766)。该基因全长2 548 bp,其中开放阅读框长2 328 bp,编码775个氨基酸,预测的蛋白分子量为84.16 kD,理论等电点为8.29。序列分析发现该基因编码的蛋白具有HMGR蛋白家族典型的HMG-CoA-reductase-classⅠ催化功能域及其他保守功能基序;序列比对和系统发育分析表明,SmHMGR与达氏按蚊Anopheles darling等长角亚目(Nematocera)昆虫HMGR的相似性最高、亲缘关系最近。SmHMGR在麦红吸浆虫滞育前的3龄早期幼虫中表达量显著升高,进入滞育后一直维持较高水平,并在滞育后静息阶段的当年12月至翌年1月达到最高。SmHMGR在蛹期表达量低于幼虫期,预蛹期表达量最低;在雌成虫中表达量显著高于在蛹和雄成虫中的表达量。【结论】SmHMGR的表达与麦红吸浆虫发育密切相关,可能在滞育诱导、维持及滞育后静息状态的维持及生殖中发挥作用,其表达量的降低可能参与了幼虫到蛹的变态。     

麦红吸浆虫热激蛋白基因SmHsp40的克隆及其在滞育进程中和极端温度胁迫下的表达

【目的】麦红吸浆虫Sitodiplosis mosellana是一种典型的专性滞育昆虫,滞育过程中经历酷暑和严寒。本研究旨在探讨麦红吸浆虫热激蛋白基因Hsp40在滞育中的作用。【方法】以麦红吸浆虫滞育前幼虫为材料,利用RT-PCR和RACE技术克隆Hsp40全长cDNA序列;利用生物信息学软件分析该基因及其编码蛋白的特性;采用实时荧光定量PCR技术分析其在滞育不同阶段幼虫及越夏滞育幼虫在短期(≤120 min)极端高温(35～50℃)和越冬滞育幼虫在短期极端低温(0～-15℃)胁迫下的表达模式。【结果】克隆获得麦红吸浆虫Hsp40基因,命名为SmHsp40(GenBank登录号:MH001167),其cDNA全长1 553 bp,开放阅读框长为1 074 bp,编码357个氨基酸,相对分子量为40.87 kD,理论等电点为5.31。推导的氨基酸序列中含有Hsp40蛋白家族典型的J结构域和HPD基序。蛋白序列相似性和系统进化关系分析表明,SmHsp40蛋白与冈比亚按蚊Anopheles gambiae等长角亚目(Nematocera)昆虫Hsp40的相似性最高、亲缘关系最近。麦红吸浆虫滞育不同时期幼虫SmHsp40表达量存在显著差异,进入滞育后表达量上调;滞育进程中,夏季7-8月和冬季12月至翌年1月表达量显著高于其他季节。与未处理的对照相比,40℃处理越夏滞育幼虫和-10℃处理越冬滞育幼虫1 h明显诱导SmHsp40的表达,但超过45℃或低于-15℃极端温度处理诱导效果均不明显。【结论】SmHsp40参与了麦红吸浆虫滞育进程,可能在滞育状态的维持及滞育期间耐热和耐寒能力的调控方面起重要作用。     

麦红吸浆虫幼虫滞育期间糖类物质变化

分析研究了麦红吸浆虫Sitodiplosis mosellana (Gehin) 幼虫滞育期间总糖、海藻糖和糖原含量的变化。结果表明： 不同滞育年限幼虫的总糖含量无显著差异。在滞育年周期中,不同时期的滞育幼虫总糖含量明显不同。幼虫老熟入土滞育以前的总糖含量明显高于入土滞育期间。入土滞育过程中,越冬期和春季的总糖含量明显高于夏季和秋季。同期的裸露幼虫与结茧幼虫的总糖含量仅在秋季有一定的差异,其他时间基本相似。结茧幼虫滞育期间的海藻糖含量一直维持较高的水平,说明其滞育期间的糖醇积累型属于海藻糖积累型;秋季与春季糖原含量的增加可能与麦红吸浆虫的滞育解除或滞育强度的变化有一定的关系。     

小麦吸浆虫保幼激素酯酶和保幼激素环氧水解酶基因的克隆及在滞育和变态发育过程中的表达动态

【目的】保幼激素(juvenile hormone, JH)在小麦吸浆虫Sitodiplosis mosellana滞育诱导及滞育后静息状态的维持中发挥着重要作用。保幼激素酯酶(hormone esterase, JHE)和保幼激素环氧水解酶(juvenile hormone epoxide hydrolase, JHEH)是调控JH滴度的重要降解酶。本研究旨在探讨JHE和JHEH在小麦吸浆虫滞育和变态发育中潜在功能。【方法】通过RT-PCR和RACE技术从小麦吸浆虫滞育前幼虫克隆JHE和JHEH全长cDNA序列;利用生物信息学软件分析其核苷酸及编码蛋白特性;采用qPCR技术分析其在小麦吸浆虫滞育不同时期(滞育前、滞育期、滞育后静息期和滞育后发育)3龄幼虫及1龄幼虫到成虫不同发育阶段(1-2龄幼虫、预蛹、初蛹、中蛹、后蛹、雌成虫和雄成虫)中的表达水平。【结果】克隆获得了cDNA全长分别为3 102和1 980 bp的小麦吸浆虫SmJHE和SmJHEH基因(GenBank登录号分别为MG876768和MG876769),其开放阅读框分别长1 740和1 371 bp,分别编码579和456个氨基酸,预测蛋白分子量分别为65.67和51.65 kD。SmJHE蛋白含有5个JHE家族特有的保守模块,SmJHEH含有催化三联体Asp228, Asp404和His431及组成阴氧离子洞的两个Tyr(Tyr299和Tyr374)和HGWP花样结构。序列比对和进化分析表明,SmJHE和SmJHEH均与双翅目(Diptera)长角亚目(Nematocera)昆虫同源蛋白氨基酸序列一致性较高,亲缘关系最近。不同滞育时期的表达模式表明,SmJHE和SmJHEH在滞育前期(1龄到滞育前的3龄幼虫早期)表达量变化不明显,进入滞育后表达量基本维持恒定,但均在滞育后静息阶段的当年12月至翌年1月最低。发育表达模式表明,幼虫恢复发育后SmJHE表达量逐渐升高,预蛹期达到最高,在雌成虫中的表达量显著低于雄成虫中的;SmJHEH表达量则在预蛹期最低,在雌成虫中最高。【结论】SmJHE和SmJHEH参与小麦吸浆虫滞育调控,其表达量的降低与滞育后静息阶段JH的累积有关;SmJHE在发育过程中表达量的升高可能参与幼虫到蛹的变态,表达量的降低可能与生殖发育有关。     

麦红吸浆虫不同滞育期四种糖代谢酶活力分析

海藻糖是麦红吸浆虫Sitodiplosis mosellana (Gehin)滞育期间储藏能量的主要物质,为弄清其在滞育期的积累机理,本文测定了麦红吸浆虫滞育前后和滞育期糖原磷酸化酶（Gpase）、己糖激酶（HK）、磷酸果糖激酶（PFK）和醛缩酶（ALD）这4种糖代谢酶活力的变化。结果表明: 麦红吸浆虫滞育前后这些糖代谢酶活力明显不同,滞育后Gpase 活力显著提高,糖酵解有关酶HK,PFK和ALD活力降低;滞育解除后,Gpase 活力降低,HK,PFK和ALD活力升高。滞育期间,4种糖代谢酶的活力均与滞育发育有关;同时Gpase 和PFK的活力也与环境温度有关,即夏、冬季高于春、秋季;同期不同滞育状态幼虫比较,裸露幼虫HK,PFK和ALD活力总是略高于结茧幼虫,Gpase则相反。滞育当年与第2年同期幼虫4种糖代谢酶的活力无显著差异。     

家蚕滞育关联山梨醇脱氢酶基因的启动子活性分析

【目的】明确家蚕Bombyx mori滞育关联基因山梨醇脱氢酶基因BmSDH(BmSDH-1,BmSDH-2a和BmSDH-2b)的转录特性。【方法】用5'RACE技术确定家蚕3个BmSDH基因的转录起始位点。利用PCR技术克隆3个BmSDH基因约1 kb及BmSDH-2a不同长度的启动子区序列,分别构建带有萤火虫荧光素酶报告基因的载体pGL3-BmSDH-P-luc,并与pRL-CMV报告质粒(含海肾荧光素酶报告基因)共转染家蚕BmN细胞,通过双荧光素酶检测系统检测BmSDH基因启动子活性;分别在BmN细胞培养基中添加昆虫保幼激素、蜕皮激素和滞育激素,通过双荧光素酶检测系统检测不同浓度激素处理对BmSDH-2a基因启动子活性的影响。【结果】BmSDH-1的转录起始位点为A(-41),BmSDH-2a的转录起始位点为C(-41),BmSDH-2b的转录起始位点为A(-40)(翻译起始位点为+1)。双荧光素酶检测结果表明,BmSDH-2a启动子活性极显著高于BmSDH-1和BmSDH-2b启动子,BmSDH-2a 355 bp长度片段的启动子活性极显著高于674 bp和1 117 bp长度片段。用不同浓度滞育激素处理BmN细胞后,BmSDH-2a的1 117 bp启动子活性随着滞育激素浓度的升高有上升的趋势,当浓度高于100 ng/mL时启动子活性有所降低但仍保持在较高水平;用保幼激素进行处理后,随着激素浓度的升高启动子活性逐渐降低;蜕皮激素处理后,0.1 ng/mL激素显著增强启动子活性,当激素浓度继续升高启动子活性逐渐降低。【结论】确定了BmSDH基因的转录起始位点。BmSDH-2a启动子活性显著高于BmSDH-1和BmSDH-2b启动子,一定浓度的蜕皮激素能显著提高BmSDH-2a启动子活性。研究结果有助于阐明BmSDH基因在家蚕滞育中的功能。     

滞育七星瓢虫的代谢适应与抗寒性评价

滞育是部分昆虫固有的适应逆境胁迫的遗传属性,七星瓢虫具有显著的滞育现象。本文以七星瓢虫雌成虫为试材,研究正常发育、滞育及滞育解除后3组处理试虫糖、脂、蛋白等关键代谢物质含量波动规律,总结滞育期间的代谢适应特点,解析其与过冷却能力的相关性,探索滞育对七星瓢虫逆境胁迫耐受力的促升效应,丰富七星瓢虫的滞育基础理论研究。利用物质干湿重差数法测定七星瓢虫的含水量;利用氯仿-甲醇(体积比为2∶1)法抽提除去自由水个体的脂肪;总糖、海藻糖、甘油、山梨醇及总蛋白的测定采用标准曲线法,利用SUN-II型智能昆虫过冷却点测定仪测定七星瓢虫的过冷却点(supercooling point,SCP)。结果表明,七星瓢虫滞育组含水量(58.11%±6.55%)显著低于正常发育组(68.49%±2.26%)和滞育解除组(65.84%±4.02%)(F=8.15,P0.01),滞育解除后含水量恢复至正常发育组水平;滞育组总糖(10.60±0.54μg/mg)、糖原(8.72±0.62μg/mg)、脂肪(173.66±19.01μg/mg)含量远远高于正常发育组和滞育解除组(F=46.57,P=0.0006;F=114.25,P0.0001;F=8.48,P0.01);滞育组总蛋白含量(49.20±3.80μg/mg)显著低于正常发育组(71.02±6.15μg/mg)和滞育解除组(69.45±4.66μg/mg)(F=46.57,P=0.0006);滞育组中海藻糖(1.31±0.27μg/mg)、甘油(1.74±0.50μg/mg)、山梨醇(9.84±3.02μg/mg)含量与正常发育组、滞育解除组无显著性差异(F=0.79,P=0.4946;F=1.33,P=0.3004;F=1.69,P=0.2387)。七星瓢虫在滞育条件下其过冷却点(-16.53℃±1.44℃)显著低于正常发育组(-14.07℃±1.33℃)和滞育解除组(-15.29℃±2.10℃)(F=13.47,P0.0001),经过滞育低温驯化后滞育解除组过冷却点较对照组有所降低。滞育诱发七星瓢虫发生显著的代谢适应,蛋白含量显著降低,抑制新陈代谢进程;糖脂含量显著升高,保障滞育维持及解除后发育的能量需求;七星瓢虫滞育属糖原积累型;滞育个体过冷却点大幅下降,耐寒性显著提升。     

麦红吸浆虫蜕皮激素受体（EcR）基因的克隆与表达分析

为了研究蜕皮激素受体（EcR）在麦红吸浆虫Sitodiplosis mosellana (Géhin)滞育活动中的作用, 利用RT PCR和RACE技术克隆得到了麦红吸浆虫蜕皮激素受体基因cDNA全长, 并通过Real-time quantitative PCR研究了其表达情况。该cDNA全长序列被命名为SmEcR （GenBank登录号： KC491135）, 其开放阅读框长1 386 bp, 编码461个氨基酸残基。其蛋白预测分子量52.90 kD, 理论等电点6.24。该蛋白与其他已报道的昆虫EcR蛋白具有很高的同源性, 其中与迟眼蕈蚊Bradysia coprophila中相应蛋白的氨基酸序列一致性高达92%。SmEcR在麦红吸浆虫不同滞育时期和不同虫态中均有表达, 且在不同滞育时期、 不同虫态中的表达量差异很大。在滞育不同时期以11月表达量最高, 12月表达量最低; 在不同虫态以麦穗幼虫中的表达量较低, 而成虫中的表达水平很高。本研究为进一步明确SmEcR在麦红吸浆虫滞育调控中的作用奠定了基础。     

Occurrence of sorbitol in Zea mays

The identity of sorbitol (d-glucitol) from maize seeds was confirmed by GC/MS of the TMSi-ether and by co-chromatography with authentic sorbitol. Sorbitol was found in seeds and silks but not in pollen or leaves. Both endosperm and embryo contained sorbitol, but endosperm accounted for most of the sorbitol recovered from intact seeds.     

Sorbitol and sorbitol phosphate in brown seaweeds

Small amounts of D-sorbitol were extracted from 15 species of brown seaweeds representing four orders, independent of seasonal effects. Sorbitol phosphate was also isolated and identified.     

Distribution of sorbitol in rosaceae

77 leaf samples representing 68 taxa of Rosaceae were investigated for the presence of sorbitol. A procedure for the quantitative estimation of sorbitol in dry plant tissues was elaborated; it made use of extraction by percolation and capillary GLC analysis of the silylated extracts. All Maloideae and Prunoideae and most Spiraeoideae were found to accumulate sorbitol. The subfamily Rosoideae was found to be heterogeneous in this respect; in most tribes sorbitol is totally lacking, but in Kerrieae, Adenostomeae and part of Dryadeae sorbitol is present in variable amounts. A clear-cut correlation between sorbitol accumulation and basic chromosome number seems to exist in Rosaceae.     

Ontogeny of sorbitol dehydrogenases in Drosophila melanogaster

It has been shown that crude extracts of Drosophila melanogaster adults contain three distinctly different enzymes which catalyze the oxidation of d-sorbitol into d-fructose. These include (1) a soluble NAD-dependent sorbitol dehydrogenase (NAD-SoDH^s), (2) a mitochondrial NAD-dependent sorbitol dehydrogenase (NAD-SoDH^m), and (3) a soluble NADP-dependent sorbitol dehydrogenase (NADP-SoDH). Developmental studies have shown that the activities of all three of these enzymes are lowest during the larval stages while highest levels are seen during or shortly prior to the adult period. With respect to NAD-SoDH^s, studies of tissue distribution in adults have shown that highest activity is associated with thoracic musculature in both sexes and with organs of the male reproductive system. The developmental profile of this enzyme reveals a significant increase in activity at between 40 and 60 hr after hatching. This time interval corresponds closely to that during which the paternally derived NAD-SoDH^s gene is expressed. An additional increase in activity is seen in male pupae at 160 hr and in female adults at 210 hr. The rapid increase in males takes place immediately following the developmental period during which the testes attach to their respective duct systems. NADP-SoDH activity is concentrated among organs of the thorax and abdomen in both sexes. Males show significantly higher levels of this enzyme during the late pupal and early adult periods. In contrast to the patterns of distribution seen for NAD-SoDH^s and NADP-SoDH, 91–92% of the total NAD-SoDH^m activity in adults is localized to the thoracic musculature. The developmental profile of this enzyme reveals a significant increase in activity during the late pupal and early adult periods, when flight muscle mitochondria are known to be proliferating and undergoing structural maturation.This work was supported in part by Grant No. GY-10830 from the National Science Foundation and a faculty research fellowship from The University of Toledo Board of Trustees.     

催化山梨醇脱水制备异山梨醇的固体酸催化剂

为提高目标产物异山梨醇的产率,考察多种固体酸催化剂催化山梨醇脱水的反应性能。结果表明:催化剂酸性与其催化性能之间有密切联系,酸性较强的H3PO4/Nb2O5催化剂显示出比其他催化剂更优异的催化性能。对磷酸负载量进行优化后,在n(P)/n(Nb)为0.8的H3PO4/Nb2O5催化剂上得到了100%的山梨醇转化率和63%的异山梨醇选择性。     

An engineered sorbitol cycle alters sugar composition, not growth, in transformed tobacco

Many efforts have been made to engineer stress tolerance by accumulating polyols. Transformants that accumulate polyols often show growth inhibition, because polyols are synthesized as a dead-end product in plants that do not naturally accumulate polyols. Here, we show a novel strategy in which a sorbitol cycle was engineered by introducing apple cDNA encoding NAD-dependent sorbitol dehydrogenase (SDH) in addition to sorbitol-6-phosphate dehydrogenase (S6PDH). Tobacco plants transformed only with S6PDH showed growth inhibition, and very few transformants were obtained. In contrast, many transgenic plants with both S6PDH and SDH were easily obtained, and their growth was normal despite their accumulation of sorbitol. Interestingly, the engineered sorbitol cycle enhanced the accumulation of sucrose instead of fructose that was expected to be increased. Sucrose, rather than fructose, was also increased in the immature fruit of tomato plants transformed with an antisense fructokinase gene in which the phosphorylation of fructose was inhibited. A common phenomenon was observed in the metabolic engineering of two different pathways, showing the presence of homeostatic regulation of fructose levels.     

A simple and inexpensive preparation of perdeuterated sorbitol for use as a biomacromolecule stabilization agent in NMR studies

Sorbitol is an excellent protein-stabilization agent, but it is typically used at high concentrations where the ¹H signals can interfere with NMR data collection and analysis. Deuteration of sorbitol can ameliorate this problem; however, perdeuterated sorbitol is not commercially available. We describe a simple and inexpensive method for preparation of perdeuterated sorbitol from perdeuterated glucose. The method is described explicitly and examples are given where the use of perdeuterated sorbitol has allowed the extraction of information, from NMR spectra, that is otherwise unobtainable.     

Effect of sorbitol addition on the physicochemical characteristics of starch–fatty acid systems

Aqueous maize starch dispersions (20%) were heated at 100 °C, in the presence of myristic, palmitic or stearic acid potassium salts as well as of sorbitol added at concentrations up to 60% (dry starch). Flow behaviour measurements at 100 °C indicated that interactions took place between the starch–fatty acid systems and sorbitol resulting in viscosity increase which was more pronounced as the sorbitol content increased. Water solubility measurements showed that a major part of sorbitol was easily extracted by excess water whereas sorption experiments revealed that the moisture uptake rate was proportional to sorbitol content of the starch systems examined. Thermomechanical studies indicated that the starch–fatty acid samples containing sorbitol up to 40% exhibited antiplasticizing behaviour. Scanning electron microscopy studies revealed that at sorbitol concentrations over 30%, free sorbitol crystals were formed on the surface of starch–fatty acid samples, whereas the percentage crystallinity as well as the crystallite size of samples were proportional to sorbitol content.     

Gluconobacter oxydans WD膜结合山梨醇脱氢酶基因在E.coli中的克隆表达

本研究构建了四株含有氧化葡萄糖酸杆菌山梨醇脱氢酶基因的重组大肠杆菌,并初步探究SldB和SldA亚基在山梨醇脱氢酶转化甘油反应中的作用。将pET28a、pETduet与PCR扩增的目的基因连接,构建单启动子调控重组质粒pET28a-sldB、pET28a-sldA、pET28a-sldBA和双启动子调控重组质粒pETduet-sldB'-sldA'。只有含pET28a-sldBA和pETduet-sldB'-sldA'的重组菌具有转化甘油的活性,表明G.oxydans WD的山梨醇脱氢酶催化甘油脱氢需要SldB和SldA亚基的共同作用。串联基因sldBA的蛋白表达结果与双启动子控制sldB和sldA基因蛋白表达结果基本相同,表明位于sldB基因末端的sldA的RBS序列可被E.coli C43的核糖体识别。     

Rapid induction of apoptosis in human gastric cancer cell lines by sorbitol

Most solid tumors, including gastric cancers, respond poorly to non-surgical treatments which are expected to induce an apoptosis-dependent involution. We hypothesize that the apoptotic machinery in solid tumors is either defective or in a suppressed condition. Overcoming the ineffective induction of apoptosis may improve the responsiveness of solid tumors to non-surgical treatments. Recently, sorbitol, a kind of hexose, has been found to be an effective inducer of apoptosis in HEp-2 cells. Therefore, it is of particular interest to examine the effect of sorbitol-treatment on gastric cancer cells. In the present study, we selected 4 gastric cancer cell lines which have been reported to exhibit different abilities in regard to apoptosis induction, and examined the effect of sorbitol-treatment on apoptosis induction. Within 3 hr after sorbitol-treatment, apoptosis was induced comparably in all cell lines examined. Cell death in MKN-1, MKN-28 or MKN-74 proceeded in a biphasic manner, while cell death in KATO-III was monophasic. The cell death partially depended on caspase activity. Treatments with sorbitol in combination with 12-O-tetradecanoylphorbol-13-acetate (TPA) markedly suppressed the apoptotic cell death, suggesting a role of protein kinase-C-dependent process. To our knowledge, this is the most rapid induction of apoptosis in human gastric cancer cells reported to date.