Molecular Mapping of a Gene for Resistance to Stripe Rust in Wheat Variety PIW138

Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most damaging diseases of common wheat （Triticum aestivum L.）. Wheat variety PIW138 introduced from Pakistan is resistant to the currently prevailing Pst race CYR32 in China. In this study, the bulked segregant analysis （BSA） method and simple sequence repeat （SSR） markers were used to map the stripe rust resistance gene in PIW138. The resistant and susceptible DNA bulks were prepared from the segregating F2 population of the cross between Thatcher, a susceptible variety as the female parent, and PIW138 as the male parent. The segregation of resistant and susceptible F2 plants inoculated with CYR32 indicated that single dominant gene determined the reactions of PIW138 line and temporarily designated as YrP138. Total 200 SSR primers were screened, and 4 SSR markers, Xwmc52, Xbarc61, Xgwm268, and Xgwm153, on chromosome 1B were found to be polymorphic between the resistant and the susceptible DNA bulks as well as their parents. Genetic linkage was tested on the segregating F2 population with 259 plants, including 196 resistant and 63 susceptible plants. All 4 SSR markers were linked to the stripe rust resistance gene in PIW138. The genetic distances of Xwmc52, Xbarc61, Xgwm268, and Xgwm153 to the resistance gene were 29.8, 6.2, 6.8, and 8.2 cM, respectively.

Cloning and Sequence Analysis of HN and F Protein Genes from a Strain of Goose Paramyxovirus

[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain.

Immunological Adjuvant Function of Aluminium Phosphate and Chicken IL-18 in NDV F Gene Vaccine

[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine （0.2 ml） containing aluminum phosphate adjuvant （90 μg）, pcDNA/F （200μg）, and pcDNA/chlL-18 （200 μg） was prepared. The 7 d old chick- ens to be tested were randomly divided into six groups （12 chickens in each group） and immunized through intramuscular injection with inactivated Newcastle disease vaccines, pcDNA/F＋pcDNA/chlL-18＋phosphate aluminum, pcDNA/F, pcDNA/F.＋pcDNA/ chlL-18, pcDNA/F＋aluminum phosphate, and physiological saline respectively; the secondary immunization was conducted with the same dose when the chickens were 21 d old. Their blood was sampled 0, 7, 14, 21, 28 d after first immunization. Anti- body titer was detected with ELISA and T cell transformation rate was measured with MIT. Experimental chicken will be challenged with 30 LD50 NDV virulence 28 d after first immunization. [Result] The survival rate of the chickens immunized with pcDNA/F＋aluminium phosphate＋pcDNA/chlL-18 achieved 8/12, higher than that of those immunized with pcDNA/F 4/12 and pcDNA/F＋pcDNA/chlL-18 （6/12）. The NDV antibody titer of the chickens immunized with pcDNA/F＋ aluminum phosphate, pcD- NA/F＋pcDNA/chlL-18 and pcDNA/F＋pcDNA/chlL-18＋aluminum phosphate is not differ- ent （P〉0.05）, but significantly lower than that of the chickens immunized with tradi- tional vaccine （P〈0.05）. The T cell transformation rate of the chickens immunized with pcDNA/F＋pcDNA/chlL-18＋aluminium phosphate was obviously higher than that of the chickens immunized with pcDNA/F （P〈0.05）. The T cell transformation rates of chickens immunized with pcDNA/F and the traditional vaccine showed no signifi- cant difference （P〉0.05）. [Conclusion] Combination of aluminium phosphate and pcD- NA/chlL-18 can significantly enhance the immune effect of NDV F gene vaccine.

上海地区鸽Ⅰ型副黏病毒F基因遗传进化及生物信息学分析

【目的】了解上海地区鸽Ⅰ型副黏病毒(PPMV-1)流行毒株F基因遗传变异情况,分析F蛋白生物信息学特征,为PPMV-1的防控提供技术支持。【方法】应用RT-PCR方法扩增分离株F基因,测序并进行遗传进化分析,并通过生物信息学在线软件对F蛋白结构功能进行预测分析。【结果】分离株F基因编码区全长为1662 bp,编码553个氨基酸,F蛋白裂解位点序列为112R-R-Q-K-R-F117,符合强毒株序列结构特征。系统进化树分析结果显示,分离株属于新城疫病毒(NDV)ClassⅡ群Ⅵ.2.1.1.2.2亚型,与疫苗株La Sota和Mukteswar处于不同进化分支,与国内分离株Pi/SH/CH/0617/2013和pigeon/Ningxia/2068/2016同属Ⅵ.2.1.1.2.2分支。生物信息学分析结果表明,F蛋白为亲水性蛋白,分子质量约为59.03 ku,理论等电点(PI)为7.87,半衰期为30 h,不稳定系数为35.06,脂肪系数为109.73。F蛋白主要分布在细胞质膜和内质网中,具有跨膜结构和信号肽,包含6个潜在的N-糖基化位点、13个半胱氨酸残基、103个O-糖基化位点,以及67个磷酸化位点。二级结构中α-螺旋、无规则卷曲、延伸链、β-转角分别占46.11%、31.46%、18.08%及4.34%,三级结构预测结果与二级结构相一致。B细胞抗原位点预测结果显示,F蛋白含有11个抗原表位(氨基酸数目≥7)。【结论】本研究分离获得的PPMV-1分离株属于NDV ClassⅡ群Ⅵ.2.1.1.2.2亚型,其F蛋白具有较好的抗原表位,可作为PPMV-1疫苗研制和抗病毒治疗的靶蛋白。研究结果为有效防控PPMV-1感染提供了一定的参考依据,同时也为进一步研发新型疫苗奠定了基础。

Relations of Budd-Chiari syndrome to prothrombin gene mutation

BACKGROUND: Budd-Chiari syndrome (BCS) is a type of disease characterized by portal hypertension and/or hy- pertension of the inferior vena cava (IVC) due to the ob- struction of the hepatic veins (HV) and/or intrahepatic IVC outlet. Being etiologically complicated and obscure, BCS can be acquired or idiopathic and several gene muta- tions may be contributable. This study was to explore whether prothrombin gene mutation (F G20210A) takes part in the pathogenesis of BCS and to investigate their cor- relativity. METHODS: In 38 proven BCS patients and 70 controls, polymerase chain reaction-restriction fragment length poly- morphism (PCR-RFLP) was used to find F G20210A mutation. To detect whether there are any mutations, four steps were taken: purification of genome DNA from whole blood, amplification of special fragment by polymerase chain reaction, digestion of the fragment via restriction en- donuclease, and analysis of results by polyacrylamide gel electrophoresis. RESULTS: F G20210A mutation was not detected in all patients and controls. CONCLUSIONS: No F G20210A mutation exists in Chi- nese patients with BCS, nor correlativity between the oc- currence of BCS and F G20210A mutation. The etiology of BCS in the Chinese needs further investigation.

蜡梅花发育相关基因CpUFO表达特性与功能分析

【目的】UFO是被子植物中首个被发现的F-box蛋白,主要参与调控花分生组织特性和花器官发育。本文通过对蜡梅CpUFO基因表达特性分析及异源表达拟南芥表型分析,初步鉴定CpUFO的功能,为阐明蜡梅花发育分子调控机制提供参考。【方法】基于前期构建的蜡梅转录组数据,克隆蜡梅CpUFO基因并对其编码蛋白进行同源比对及进化树分析。根据qRT-PCR分析CpUFO基因在不同组织、器官及全年花发育各阶段中的相对表达量,比较35S::CpUFO转基因拟南芥株系及Col-0野生型表型差异,并通过qRT-PCR检测过表达拟南芥株系内源开花相关基因的表达特性,分析过表达CpUFO对拟南芥开花时间以及花器官发育的影响。【结果】获得的CpUFO基因cDNA序列为1665 bp,编码403个氨基酸,含有保守的F-box结构域。CpUFO在外花被片中表达量相对最高,其次是内花被片,在果、茎、叶、腋芽及雌雄蕊中表达水平较低;而在全年花芽中的表达量分析结果显示,低温打破休眠后的露瓣和始花以及盛开期花芽中的表达量显著高于其他时期。与野生型拟南芥相比,35S::CpUFO转基因株系莲座叶数目减少,开花提前,且拟南芥内源基因TFL1表达量下调,成花关键基因FUL和AP1的表达量上调。【结论】CpUFO在不同组织、器官及花发育各阶段均有表达,不同组织中在外花被片中表达量最高,全年花芽中打破休眠的花蕾开放阶段表达量最高。同时,过表达CpUFO会促进拟南芥早花并影响其花器官发育。

莲草直胸跳甲短神经肽F受体基因sNPFR的克隆及表达分析

为明确莲草直胸跳甲(Agasicles hygrophila)短神经肽F受体AhsNPFR功能及其表达特点,为探索莲草直胸跳甲生防作用奠定理论基础,利用PCR技术克隆鉴定莲草直胸跳甲AhsNPFR基因并进行生物信息学分析,通过实时荧光定量PCR技术分析其在莲草直胸跳甲不同发育时期和组织中的时空表达谱。结果表明,克隆获得基因AhsNPFR全长1 669 bp,开放阅读框1 257 bp,编码418个氨基酸;预测其蛋白质分子质量为48.11 ku,理论等电点为8.21,AhsNPFR具有7个典型保守跨膜结构域,属于GPCRs家族,系统进化树分析表明,其与玉米根萤叶甲Diabrotica virgifera virgifera sNPFR亲缘关系最近。AhsNPFR基因在不同发育阶段均有表达,在1龄幼虫中的表达量最高,是卵中表达量的9.06倍;在卵中表达量最低;雌成虫的表达量显著高于雄成虫。AhsNPFR基因在不同组织中均有表达,在3龄幼虫后肠中显著高表达,是脂肪体表达量的12.21倍;在雌雄成虫后肠显著高表达,并在所有组织中均没有雌雄表达差异。

Construction of Recombinant Expression Plasmids Containing H and F Protein Genes of Canine Distemper Virus Isolated from a Mink and Their Expression in Prokaryotic Cells

[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well as to study the reactogenieity of the expressed products. [ Method ] RT-PCR amplification was used to obtain H and F protein genes; TA cloning and subclonlng techniques were used to construct the cloning plasmids（pMD-18T-H and pMD-18T-F） and recombinant expression plasmids（pET28a-H and pET28a-F） ; SDS-PAGE and Western-blotting were adopted to verify whether the target proteins were successfully expressed. [ Result] The recombinant expression plasmids pET28a-H and pET28a-F containing H and F protein genes of Canine distemper virus isolated from a mink were successfully constructed, and both the expressed H and F proteins with respectively relative molecular mass of 31 400 and 38 200 produced positive reac- tion with the CDV standard positive serum. [ Conclusion] The H and F proteins expressed in prokaryotic cells were the same with the natural ones in terms of reac- togenicity, which can be utilized for diagnosis of a CDV＇s infection or for an epidemiological investigation. Meanwhile, they also provide a basis for developing ge- netically engineered subunit vaccines.

Genetic Analysis and Molecular Mapping of a Stripe Rust Resistance Gene YrH9014 in Wheat Line H9014-14-4-6-1

Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most widespread and destructive wheat diseases in many wheat-growing regions of the world. The winter wheat translocation line H9014-14-4-6-1 has all stage resistance. To identify stripe rust resistance genes, the segregating populations were developed from the cross between H9014-14-4-6-1 and Mingxian 169 （a wheat cultivar susceptible to all Pst races identified in China）. The seedlings of the parents and F1 plants, Fz, F3 and BC1 generations were tested with Pst races under controlled greenhouse conditions. Two genes for resistance to stripe rust were identified, one dominant gene conferred resistance to SUN11-4, temporarily designated YrH9014 and the other recessive gene conferred resistance to CYR33. The bulked segregant analysis and simple sequence repeat （SSR） markers were used to identify polymorphic markers associated with YrH9014. Seven polymorphic SSR markers were used to genotype the F2 population inoculated with SUN11-4. A linkage map was constructed according to the genotypes of seven SSR markers and resistance gene. The molecular map spanned 24.3 cM, and the genetic distance of the two closest markers Xbarc13 and Xbarc55 to gene locus was 1.4 and 3.6 cM, respectively. Based on the position of SSR marker, the resistance gene YrH9014 was located on chromosome arm 2BS. Amplification of a set of nulli-tetrasomic Chinese Spring lines with SSR marker Xbarc13 indicated that YrH9014 was located on chromosome 2B. Based on chromosomal location, the reaction patterns and pedigree analysis, YrH9014 should be a novel resistance gene to stripe rust. This new gene and flanking markers got from this study should be useful for marker-assisted selection （MAS） in breeding programs for stripe rust.

^(18)F-FDG PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者KRAS基因突变的评估价值

目的探讨^(18)F-氟代脱氧葡萄糖(^(18)F-FDG)PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者鼠类肉瘤病毒癌基因(KRAS)基因突变的评估价值。方法回顾性分析64例晚期结直肠癌患者的临床资料。所有患者在治疗前均接受^(18)F-FDG PET/CT检查。分析最大标准化摄取值(SUV_(max))、不同阈值下的代谢肿瘤体积(MTV)和病变总糖酵解(TLG)、临床病理特征与患者KRAS基因突变状态之间的关系。通过多因素Logistic回归模型分析与患者KRAS基因突变相关的因素。采用受试者工作特征曲线分析^(18)F-FDG PET/CT代谢参数、临床病理特征及二者联合评估患者KRAS基因突变的效能。基于^(18)F-FDG PET/CT代谢参数和临床病理特征构建列线图模型。结果单因素和多因素Logistic回归分析结果显示,SUV_(max)≥19.55、MTV50%≥7.95、肿瘤组织中/高分化与患者KRAS基因突变有关。SUV_(max)、MTV50%和肿瘤组织分化程度评估患者KRAS基因突变的曲线下面积(AUC)分别为0.653、0.625和0.621,三者联合评估的AUC为0.800,均高于单个指标的AUC(P<0.05)。基于SUV_(max)、MTV50%和肿瘤组织分化程度构建的列线图模型的一致性指数为0.800,校准曲线与参考线基本拟合。结论^(18)F-FDG PET/CT的代谢参数SUV_(max)、MTV50%与晚期结直肠癌患者KRAS基因突变密切相关,联合肿瘤组织分化程度所构建的列线图模型对患者KRAS基因突变具有较高的评估价值。

一体化^(18)F-FET PET/MR术前评估成人胶质瘤患者MGMT基因启动子甲基化状态

目的 旨在研究一体化^(18)F-氟乙基酪氨酸(^(18)F-fluoroethyltyrosine, ^(18)F-FET)正电子发射断层成像(positron emission tomography, PET)/MR对成人胶质瘤的O6-甲基鸟嘌呤DNA甲基转移酶(O6-methylguanine DNA methyltransferase, MGMT)基因启动子甲基化状态的鉴别能力。材料与方法 回顾性分析未经活检或治疗的胶质瘤患者资料16例,均完成一体化PET/MR扫描,包括^(18)F-FET PET、常规MRI及体素内不相干运动(intravoxel incoherent motion, IVIM)成像。以靶本比(target-background ratio, TBR)=1.6为阈值对PET图像进行感兴趣体积(volume of interest, VOI)分割,通过刚性配准获得肿瘤VOI对应的IVIM图及其参数表观扩散系数(apparent diffusion coefficient, ADC)、真实扩散系数(true diffusion coefficient, D)、伪扩散系数(pseudo diffusion coefficient, D^(*))、灌注分数(perfusion fraction, f)、分布扩散系数(distributed diffusion coefficient, DDC)和异质性指数(heterogeneity index, α),使用pyradiomics对各参数进行特征提取,获得对应的一阶灰度直方图特征。计算每个IVIM参数图对应的特征值与^(18)F-FET PET参数的关系,并利用组间比较和受试者工作特征(receive operating characteristic, ROC)曲线分析探究PET和IVIM参数对MGMT启动子甲基化的区分能力。结果 IVIM-α的两个特征值90分位值(r=0.526,P<0.05)和最大值(r=0.520, P<0.05)与PET参数平均标准摄取值(meanstandarduptakevalue,SUVmean)呈正相关;IVIM-α和SUVmean在MGMT启动子甲基化状态阳性和阴性两组之间的差异有统计学意义(P<0.05),阳性组的IVIM-α均值和SUVmean显著高于阴性组;融合IVIM-α均值和SUVmean对MGMT启动子甲基化状态进行区分的曲线下面积(area under the curve, AUC)为0.77。结论 基于一体化PET/MR的^(18)F-FET PET和IVIM参数能够有效预测胶质瘤的MGMT启动子甲基化状态。

6株鸽源新城疫病毒广东分离株F、HN基因的克隆及遗传进化分析

为研究广东省鸽源新城疫病毒(NDV)的遗传进化规律,试验于2021—2022年从广东6家疑似感染NDV的鸽场采集病鸽临床样品,进行RT-PCR诊断、病毒分离鉴定和动物回归试验,并对分离毒株进行生物学毒力测定以及F基因、HN基因的同源性和遗传进化分析。结果显示:共分离到6株鸽源NDV,对幼鸽进行分组攻毒14 d后,发病率均为100%,死亡率为60%~80%;6株毒株的F蛋白裂解位点氨基酸序列为112R-R-Q-K-R-F117/112R-R-R-K-R-F117,具有NDV强毒株的典型分子特征,和分离毒株ICPI的测定结果共同表明6株毒株均为强毒株;6株毒株均为NDV的ClassⅡ基因Ⅵ.2.1.1.2.2型;与传统疫苗株La Sota的同源性较低,且F、HN蛋白氨基酸出现多处位点变异。研究表明,广东省鸽源NDV流行以Class Ⅱ基因Ⅵ.2.1.1.2.2型为主,常用的疫苗株La Sota可能对广东地区流行的鸽源NDV保护效果不佳。

Inheritance and Molecular Mapping of Stripe Rust Resistance Gene Yr88375 in Chinese Wheat Line Zhongliang 88375

Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient strategies for breeding cultivars resistant to stripe rust. Zhongliang 88375, a common wheat line, is highly resistant to all three rusts of wheat in China. The gene conferring rust disease was deduced originating from Elytrigia intermedium. Genetic analysis of Zhongliang 88375 indicated that the resistance to PST race CYR31 was controlled by a single dominant gene, temporarily designated as Yr88375. To molecular map Yr88375, a F2 segregating population consisting of 163 individuals was constructed on the basis of the hybridization between Zhongliang 88375 and a susceptible wheat line Mingxian 169; 320 SSR primer pairs were used for analyzing the genetic linkage relation. Six SSR markers, Xgwm335, Xwmc289, Xwmc810, Xgdmll6, Xbarc59, and Xwmc783, are linked to Yr88375 as they were all located on chromosome 5BL Yr88375 was also located on that chromosome arm, closely linked to Xgdmll6 and Xwmc810 with genetic distances of 3.1 and 3.9 cM, respectively. The furthest marker Xwmc783 was 13.5 cM to Yr88375. Hence, pedigree analysis of Zhongliang 88375 combined with SSR markers supports the conclusion that the highly resistance gene Yr88375 derived from Elytrigia intermedium is a novel gene for resistance to stripe rust in wheat. It could play an important role in wheat breeding programs for stripe rust resistance.

The SNPs C.513A>T in the MHC B-F gene and rs15001532 in the SPOCK1 gene are associated with Salmonella pullorum disease resistance in chickens

supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）

Molecular Mapping of a Stripe Rust Resistance Gene YrH9020a Transferred from Psathyrostachys huashanica Keng on Wheat Chromosome 6D

Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.

Genetics and Molecular Mapping of Stripe Rust Resistance Gene YrShan515 in Chinese Wheat Cultivar Shan 515

Stripe rust is one of the most important wheat diseases worldwide. To identify new resistance genes is significant in wheat breeding. In this study, stripe rust resistance of a Chinese cultivar Shan 515 was tested with Chinese predominant races of P. striiformis f. sp. tritici in the seedling stage, and genetic analysis and simple sequence repeats （SSR） technique were used to identify the inheritance model of seedling stripe rust resistance in cultivar Shan 515 and to mark the sites of resistance gene（s） on chromosome. The genetic analysis indicated that the resistance of Shan 515 against Su11-4 was conferred by a single dominant gene, which was temporarily designated as YrShan515. Using bulked segregant analysis （BSA） and SSR markers, 12 SSR markers （Xwmc335, Xwmc696, Xwmc476, Xbarc267, Xgwm333, Xwmc653, Xwmc396, Xgwm213, Xgwm112, Xgwm274, Xcfd22, Xgwm131, and Xwmc517） located on wheat chromosome 7BL were linked to YrShan515 with genetic distance ranging from 3 to 24 cM. Based on the previously published genetic map and Chinese Spring nulli-tetrasomic analysis, YrShan515 was located on wheat chromosome 7BL. Polymorphism of wheat cultivars collected from Huanghuai wheat grown regions were screened with two markers, Xwmc653 and Xbarc267, and all of these wheat cultivars tested did not present the polymorphic bands as Shan 515 did. Therefore, it suggested that YrShan515 might be a allele of the available yellow rust resistance gene. The mapping of the new resistance gene in Shan 515 is useful for wheat breeding and diversification of resistance genes against stripe rust in commercial wheat cultivars in China.

HbA、HbA2及HbF与地中海贫血基因检测联合诊断地中海贫血的临床意义

目的本研究将血红蛋白A(HbA)、血红蛋白A_(2)(HbA_(2))及血红蛋白F(HbF)与地中海贫血基因检测联合诊断应用于地中海贫血的诊断中,并对该方式的诊断效能进行分析。方法70例疑似地中海贫血患者,随机分为研究组和对照组,每组35例。同期纳入在本院接受健康体检的50例正常人作为健康组。对照组和健康组进行血红蛋白电泳检测(HbA、HbA_(2)、HbF),研究组实施血红蛋白电泳检测和地中海贫血基因检测。对比研究组和对照组患者的地中海贫血检出率,并对比α-地中海贫血患者、β-地中海贫血患者以及健康组的HbA、HbA_(2)及HbF水平,分析HbA、HbA_(2)、HbF联合地中海贫血基因检测诊断地中海贫血的受试者工作特征曲线(ROC曲线)。结果研究组的α-地中海贫血检出率为34.29%、β-地中海贫血检出率为54.29%,对照组的α-地中海贫血检出率为31.43%、β-地中海贫血检出率为37.14%,研究组地中海贫血总检出率(88.57%)高于对照组(68.57%),差异有统计学意义(P<0.05)。α-地中海贫血患者、β-地中海贫血患者、健康组的HbA水平对比,差异无统计学意义(P>0.05);α-地中海贫血患者、β-地中海贫血患者、健康组的HbF、HbA_(2)水平对比,差异有统计学意义(P<0.05)。其中β-地中海贫血患者的HbF水平(5.87±1.56)%、HbA_(2)水平(5.62±1.67)%高于α-地中海贫血患者的(1.67±0.52)%、(1.51±0.34)%以及健康组的(1.21±0.19)%、(2.81±0.92)%;α-地中海贫血患者的HbA_(2)水平低于健康组,而HbF水平高于健康组,差异有统计学意义(P<0.05)。以HbA、HbA_(2)、HbF与地中海贫血基因检测为检验变量,设置1为患病、0为未患病作为效应变量绘制ROC曲线:其中HbA、HbA_(2)、HbF诊断的曲线下面积(AUC)值为0.703,95%置信区间(CI)=(0.554,0.852),灵敏度为67.3%,特异度为73.3%;HbA、HbA_(2)、HbF联合地中海贫血基因检测诊断的AUC值为0.852,95%CI=(0.736,0.967),灵敏度为83.6%,特异度为86.7%。结论在对地中海贫血进行诊断时采取HbA、HbA_(2)、HbF联合地中海贫血基因检测,具有较高的诊断效能,并且能够有效鉴别地中海贫血的疾病类型,在后续开展的相关治疗中具有重要意义。

丹参FBXL基因家族的鉴定和表达模式分析

目的基于基因组数据鉴定丹参F-box-LRR(FBXL)基因家族,并对其进行生物信息学与表达模式分析,为进一步深入阐明其基因功能提供依据。方法从丹参基因组数据库中鉴定出SmFBXL基因,运用生物信息学方法及在线工具分析其基因结构特征,启动子顺式作用元件,编码的蛋白理化性质、系统进化、组织表达等。结果从丹参的基因组中共鉴定出104个SmFBXL基因(SmFBXL1~SmFBXL104),不均等分布于8条染色体上,上游启动子含有与植物抗逆、生长发育和激素应答等相关的顺式作用元件。构建丹参、拟南芥和大豆的FBXL家族成员的系统发育树,将104个SmFBXL基因分为7个亚族。通过同源进化分析,猜测SmFBXL36可能参与防御病原菌入侵,SmFBXL86、SmFBXL79可能在调控丹参侧根生长中起重要作用,SmFBXL11、SmFBXL40可能调节下胚轴生长。转录组数据显示SmFBXL基因在丹参不同组织中差异表达,其中13个SmFBXL基因在根和叶中的表达水平较高,可作为后续研究SmFBXL基因家族的候选基因。结论研究结果为进一步解析SmFBXL基因在丹参逆境响应及次生代谢产物生物合成中的调控机制提供了参考。

Association Analysis of SP-SNPs and Avirulence Genes in Puccinia striiformis f. sp. tritici, the Wheat Stripe Rust Pathogen

Puccinia striiformis f. sp. tritici (Pst) is one of the pathogenic fungi on wheat, caused stripe rust that is a great threat for wheat production all over the world. Intensive efforts have been made to study genetics of wheat resistance to this disease, but few on avirulence of the pathogen due mainly to the nature of obligate biotrophism and the lack of systems for studying its genetics and molecular manipulations. To overcome these limitations, a natural Pst population comprising 352 isolates representative of a diverse virulence spectrum was genotyped using 97 secreted protein-single nucleotide polymorphism (SP-SNP) markers to identify candidate avirulence genes using association analysis. Among avirulence genes corresponding to 19 resistance genes, significantly associated SP-SNP markers were detected for avirulence genes AvYr1, AvYr2, AvYr6, AvYr7, AvYr8, AvYr44, AvYrExp2, AvYrSP, and AvYrTye. These results indicate that association analysis can be used to identify markers for avirulence genes. This study has laid the foundation for developing more SP-SNPs for mapping avirulence genes using segregating populations that can be generated through sexual reproduction on alternate hosts of the pathogen.

新城疫病毒F_(48)E_9株M NP F和HN基因的真核表达

将新城疫病毒(NDV)F48E9株的M、NP、F和HN基因克隆到真核细胞表达载体 pCAGG上,经酶切、PCR鉴定和序列分析,分别筛选出了含有M、NP、F和HN基因的重组质粒, 命名为pCAGG-M、pCAGG-NP、pCAGG-F和pCAGG-HN。纯化后的重组质粒通过脂质体单独转染HeLa细胞,经间接免疫荧光试验检测到了M、NP、F和HN蛋白的表达。pCAGG-M、 pCAGG-NP、pCAGG-F和pCAGG-HN重组质粒组合共转染HeLa细胞48 h后,可以观察到明显的细胞融合现象。试验结果表明,NDV F48E9株的M、NP、F和HN蛋白均在HeLa细胞内得到了成功表达,同时证明在该表达系统中F蛋白不足以诱导细胞融合。