谷氨酰胺对斜带石斑鱼GF-1细胞中C-Myc蛋白的表达与神经坏死病毒复制的影响

为探讨C-Myc表达、谷氨酰胺代谢和神经坏死病毒复制三者之间的关系,本研究首先克隆了斜带石斑鱼鳍条细胞(GF-1)中的C-Myc基因(GF-1-C-Myc),结果显示GF-1-CMyc基因cDNA全长814 bp,开放阅读框(ORF)为285 bp,编码95个氨基酸(aa),有亮氨酸拉链结构域与螺旋-环-螺旋(HLH)结构域。实验表达和纯化了GF-1-C-Myc蛋白,并制备其多克隆抗体。采用实时定量PCR技术(qRT-PCR)与免疫印迹法(WB)检测了GF-1-C-Myc基因的表达和神经坏死病毒的复制。结果显示,缺乏谷氨酰胺会同时抑制GF-1-C-Myc基因的表达和神经坏死病毒(NNV)的复制,添加谷氨酰胺可同时促进GF-1-C-Myc的表达和NNV的复制;此外,NNV感染可上调GF-1-C-Myc基因的表达,并显著消耗GF-1细胞培养液中的谷氨酰胺。研究表明,GF-1-C-Myc基因可调控宿主谷氨酰胺代谢,从而有利于神经坏死病毒的复制。本结果为防控NNV的感染提供了参考。     

Verrucosamide,a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD₅₀ of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD₅₀ of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.     

罗氏沼虾细胞色素P450家族CYP302a1基因克隆及其在蜕皮周期中的表达

蜕皮是甲壳动物重要的生理活动,与其蜕皮激素的合成密切相关,细胞色素P450(CYP)302a1是甲壳动物蜕皮激素合成通路中的关键酶之一。本研究克隆了罗氏沼虾CYP302a1基因(Mr-CYP302a1),cDNA全长1859 bp,开放阅读框(ORF)为1629 bp,编码543个氨基酸(aa),分子量大小为61.09 ku,等电点为8.42。氨基酸序列分析显示CYP302a1基因的保守结构域含有5个P450基因家族特征保守区域:heme-binding、helix-K、helixC、helix-I及PERF。系统进化分析结果显示Mr-CYP302a1首先与绿虾CYP302a1聚为一支,然后与凡纳滨对虾及三疣梭子蟹等十足目甲壳动物的CYP302a1聚为一支,与甲壳动物的亲缘关系最近。实时荧光定量PCR(qRT-PCR)检测表明Mr-CYP302a1在罗氏沼虾的多个组织中均有表达,其中在Y器官中的表达量最高,性腺中次之。同时研究发现,MrCYP302a1基因在罗氏沼虾的蜕皮后期(A期和B期)表达量很低,蜕皮间期(C期)表达量开始上升,在蜕皮前期D1亚期达到峰值。对Mr-CYP302a1进行蛋白表达及多克隆抗体制备,蛋白印迹法(Western blot,WB)检测表明Mr-CYP302a1蛋白在罗氏沼虾Y器官中的表达量最高,在蜕皮过程中的蜕皮前期D1亚期达到峰值。综上所述,该基因在罗氏沼虾的蜕皮过程中扮演着十分重要的角色。     

稻曲病菌Six1类效应蛋白UvSix1-1的功能研究

 稻曲病菌在侵染过程中会分泌大量的效应蛋白来帮助其侵染。本研究鉴定到一个稻曲病菌效应蛋白UvSix1-1,同源序列比对及系统发育分析发现其具有Six1蛋白保守的氨基酸位点,并与多个病原菌中的Six1蛋白具有同源性。进一步研究发现UvSix1-1可以抑制Bax、XEG1和INF1引起的烟草叶片细胞坏死,并且其预测的信号肽区域具有分泌功能,在烟草上瞬时表达UvSix1-1可以促进辣椒疫霉的定殖。对不同稻曲菌株中的序列进行分析,没有发现序列多态性,说明该基因非常保守。以上研究结果表明,UvSix1-1在病原菌侵染过程中发挥作用,研究结果为稻曲病菌效应蛋白的研究提供参考。     

利用CRISPR/Cas9敲除葡萄VviPDS1基因的研究

选择葡萄八氢番茄红素脱氢酶(Phytoene desaturase)基因VviPDS1为靶标,利用CRISPR/Cas9系统构建基因敲除载体,瞬时转化葡萄叶片原生质体,检测到不同类型的突变。通过农杆菌介导转化‘无核白’葡萄胚性愈伤组织,筛选获得卡那霉素抗性植株71株。经PCR鉴定,其中53株为阳性植株,阳性率为74.64%。测序结果表明,共有20株在靶点发生不同类型的突变,编辑效率为37.74%;其中9株产生了双等位基因突变。对其进行氨基酸序列预测,在第202位氨基酸之后发生了不同程度的变异。利用CRISPR/Cas9系统敲除VviPDS1获得的突变体植株呈现整体矮化,其叶片出现不同程度白化。表明CRISPR/Cas9系统可以通过细胞中的瞬时或稳定表达进行基因编辑,可以实现在葡萄编辑植株中产生纯合敲除。     

樱桃番茄抗青枯病砧木新品种海茄砧1 号的选育

海茄砧1 号是以野生茄子自交系HZ09-5 为母本，以从越南引进的茄子品种自交系HZ10-2 为父本育成的高抗青枯 病樱桃番茄砧木品种。植株生长势较强，第1 雌花节位约为第7 节，花为紫色，果实卵圆形，青熟果淡绿紫色或淡绿色，果 萼绿色，果长7～9 cm，果宽4～6 cm，单果质量为50～60 g，种子千粒重为3.2 g 左右，种皮浅黄色，高抗青枯病。海茄砧 1 号与樱桃番茄的嫁接亲和性好，共生性强，嫁接苗成活率高。嫁接后的樱桃番茄果萼开展，果面有光泽，可明显改善果实 VC、可溶性固形物和可滴定酸含量等品质，提高产量。适宜华南地区樱桃番茄的嫁接栽培。     

具有解析式位置正解的2T1R并联机构运动性能分析

求解具有解析式位置正解且部分运动解耦的并联机构,有利于后续的误差分析、动力学分析、运动轨迹规划与控制等。基于方位特征方程(POC)的并联机构设计理论与方法,设计了两种具有解析式位置正解且部分运动解耦的2T1R并联机构,并对这两种机构进行了方位特征、自由度及耦合度等主要拓扑性能分析;提出基于拓扑特征的运动学建模与求解方法,并据此求解了两种机构的解析式位置正解;基于导出的位置反解,分析了两种机构工作空间、奇异位形、动平台的速度与加速度变化规律。最后比较了两种机构的运动性能,选择了优选机型。为优选机型的动力学分析与样机研制提供了理论基础。     

串番茄新品种红星302 的选育

红星302 是以抗病、耐低温弱光材料01-61 为母本，以01-23 为父本育成的串番茄一代杂种，无限生长类型，果实串收，果色亮红，果实椭圆形，酸甜适口，品质佳，VC 含量为364 mg · kg^-1，可滴定酸0.44%，可溶性糖4.25%，平均单果质量30.79 g，硬度为11.40×10⁵ Pa。单株结果数约为53个，产量4 900 kg ·（ 667 m²）^-1 左右，高抗蕨叶病毒病、青枯病，适宜河北地区冬季日光温室和早春保护地栽培。     

甜樱桃花芽不同发育时期内参基因的筛选与验证

为了筛选甜樱桃花芽不同发育时期均稳定表达的内参基因,以甜樱桃桑提娜和黔樱一号不同发育时期花芽为材料,通过qRT-PCR技术检测28S rRNA、EF1-a1、EF1-a2、UBC、RPL13、18S rRNA、RSP3、CYP40、ACT2和α-TUB3等10个常用看家基因的表达水平,并利用GeNorm、NormFinder和BestKeeper综合评价其表达稳定性。结果表明,EF-1a2和RSP3在所有样品中稳定性最好。分别以EF-1a2、RSP3及EF-1a2+RSP3作为内参基因检测不同发育时期花芽生长素运输载体AUX1基因及生长素响应因子ARF基因的表达模式,该2个基因在不同内参基因标定下表达模式相同。表明EF-1a2、RSP3及EF-1a2+RSP3可作为甜樱桃花芽不同发育时期的内参基因。     

Effect of pidotimod on renal function and inflammatory response in rat IgA nephropathy model

AIM:To explore the effect of pidotimod on the renal function in IgA nephropathy (IgAN) rat model, and to further study whether this effect is related to the inhibition of inflammatory response. METHODS:The SD rats (n=36) were randomly divided into control group, IgAN model group, IgAN with prednisone treatment group and IgAN with pidotimod treatment group, with 9 rats in each group. The IgAN model was induced by consecutive oral administration of bovine gamma globulin (BGG) for 8 weeks followed by injection of BGG through tail vein for 3 d. After the IgAN model was established, the drug was continuously used for 4 weeks. At the end of the treatment, the urine protein, serum creatinine and blood urea nitrogen were examined by an automated analyzer. IgA deposition in the renal tissues was observed by immunofluorescence staining. The mRNA expression levels of renal fibrosis markers transforming growth factor-β1 (TGF-β1) and fibronectin 1 in the renal tissues were detected by RT-qPCR. The mRNA and protein levels of pro-inflammatory cytokines interleukin-1β (IL-1β) and IL-6 in the renal tissues were determined by RT-qPCR and Western blot, respectively. RESULTS:No significant difference of the body weight was observed in different groups. Compared with control group, the content of urine protein, serum creatinine and blood urea nitrogen were significantly increased (P<0.01), whereas those were reversed by pidotimod treatment. The results of immunofluorescence staining showed that pidotimod inhibited IgA deposition in the IgAN rats. Pitomod treatment inhibited the mRNA expression levels of renal fibrosis markers TGF-β1 and fibronectin 1, and the mRNA and protein levels of pro-inflammatory cytokines IL-1β and IL-6 in the renal tissues of IgAN rats. CONCLUSION:Pidotimod alleviates IgAN progression in rats by inhibition of inflammatory response.