郑州地区汉族人群Gc亚型遗传多态性研究

木文用超薄聚丙烯酸胺凝胶等电聚焦结合磺基水杨酸沉淀方法,对郑州地区311名汉族无关个休的血清样本进行了Gc亚型分型。除见到了6种常见的表现型外,还检测到4种罕见变异型。其基因频率为：Gc^1F=0.4035, Gc^1s＝0.2669,Gc²=0.3200,Gc^v=0.0096,经用Hardy-Weingberg定律检验,观测值和期望值吻合良好。     

用固相pH梯度等电聚焦分析Gc蛋白亚型

用固相pH梯度等电聚焦技术及免疫固定技术对201名北京地区无关汉族群体的人血清Gc蛋白亚型进行了分型鉴定及基因频率的调查.Gc^1F为0.3698.Gc^1S为0.2812.Gc²为0.3490.观察值与期望值吻合良好(Σx²=1.057,P＞0.70)     

用Phast System自动电泳仪快速测定血清CK-MM亚型及初步临床应用

本文介绍了在Phast System全自动电泳仪上用等电聚焦法对血清CK-MM亚型的快速测定。灵敏度高,分辨率好,用1小时即可完成测定,特别适合临床需要。30例正常人血中CK-MM₁含量为44.3％,CK-MM₂为38.9％,CK-MM₃含量最低为16.7％,MM₃/MM₁比值小于1。而12例患者血中CK-MM亚型有明显变化,初步证实本法对早期诊断急性心肌梗塞(AMI)和溶栓治疗后判断是否出现再灌注有较高的临床价值。     

微量全血快速检测乙肝病毒基因亚型

选用特殊血液标本裂解液,可有效除去血液中对Taq酶有抑制作用的物质血叶啉,微量指尖全血标本即可满足检测需要,可取代血清检测HBV基因亚型．     

一种简单、快速、微量聚丙烯酰胺凝胶等电聚焦电泳方法

Bio-Rad公司的微量聚丙烯酰胺凝胶等电聚焦电泳(ModelⅢ Mini IEF Cell),不需要使用电极缓冲液,聚焦电泳时间仅用1.5小时,效果甚佳,但要使用该公司专有的凝胶支持薄膜,难于推广应用,我们经过试验,建立了一种简单、快速的微量凝胶等电聚焦电泳方法,40min完成等电聚焦过程,采用快速染色、脱色和干燥,将电泳图谱制成透明的薄膜,整个过程可以     

上海地区胃癌病人运铁蛋白(Tf)、组特异性成分(Gc)、α_1-抗胰蛋白酶(α_1-AT)亚型分布的研究

用薄层聚丙烯酰胺凝胶等电聚焦电泳分析了上海地区202例汉族胃癌病人及202例正常对照组的运铁聋白(Tf)和α_1-抗胰蛋白酶(α_1-AT)亚型的分布,发现胃癌组Tfc_1c_1纯合子频率(0.3713)和Tfc_1基因频率(0.5718)显著低于对照组(分别为0.5149和0.6782),均为p<0.01,胃癌组Tfc_2c_2纯合子频率(0.2228)和Tfc_2基因频率(0.4019)显著高于对照组(分别为0.1436,p<0.05和0.2970,p<0.01),胃癌组和对照组的α_1-抗胰蛋白酶亚型分布无显著差异。用薄层聚丙烯酰胺等电聚焦结合免疫固定分析了上海地区200例汉族胃癌病人和200例正常对照组的组特异性成分(Gc)亚型分布,发现胃癌组Go1F表型频率(0.22)和Gc1F基因频率(0.4375)均显著高于正常对照组(分别为0.14和0.3600,均为p<0.05)。     

超高分辨率毛细管等电聚焦—电喷雾质谱联用方法观察蛋白质亚型

在线的毛细管等电聚焦 电喷雾质谱联用 ,作为一种二维的分离系统 ,对毛细管等电聚焦过程中形成的蛋白质亚型进行了分析。这种分析系统通过使用中性的涂层毛细管 (80cm长 )、动态的毛细管位置调整方法和鞘流液接口得以建立。蛋白质首先在毛细管等电聚焦过程中根据它们等电点的差异得到分离 ,然后被电喷雾质谱鉴定。已聚焦好的蛋白质区带通过结合阴极移动和重力移动的方法从毛细管中流出而进入质谱仪。由于在此特定情况下这种方法具有极高的分辨率 ,有三种血红蛋白A和镰刀型血红蛋白的亚型 (具有几乎相同的电荷分布和分子质量 ,但它们的等电点差异在 0 .0 4到 0 .0 8之间 )和两种乳球蛋白A的亚型 (等电点差异为 0 .6 )被检测到。这些蛋白质亚型的等电点、相对含量和分子质量都通过毛细管等电聚焦 电喷雾质谱联用方法同时得到了确定。     

用等电聚焦技术对两种斑鸠卵清蛋白的比较

用聚丙烯酰胺凝胶等电聚焦技术进行了珠颈斑鸠(Strptopelia chinensis chinensis)和山斑鸩(S.Orientalis orietalis)的卵清蛋白比较分析。结果表明.在pH3.5—10的梯度内,山斑鸠为21条带。珠颈斑鸠为18条带。其中主要区带的pI在4.5—6.0之间.其谱带较近似。在蛋白质分子水平上,两者是亲缘关系十分相近的两个种。     

Gc subtypes in the middle east: Report on an Arab Moslem population from Israel

Gc subtypes were determined by isoelectrofocusing and immunofixation on 342 blood samples from an Arab Moslem population in Israel. Observed allele frequencies were: Gc^1F 0.2120, Gc^1S 0.6023, and Gc² 0.1857. Those are similar to formerly reported frequency data for other Middle Eastern populations. A discriminant analysis, performed on data from 35 populations, resulted in a satisfactory classification of population groups related through geographic and racial origin.     

Gc and transferrin isoelectrofocusing subtypes among Brazilian Indians

A total of 397 persons living in six villages of three Brazilian Indian tribes were studied in relation to the Gc subtypes. The corresponding gene frequencies are more similar between the Gorotire and Caingang than between the Gorotire and the Krahó, despite the considerable geographical distance that separates the villages of these two first tribes and their lignuistic differentiation. An uncommon variant pattern (1C7) was observed in eight Gorotire Indians; it had been described for the first time in a Tibetan sample, furnishing additional evidence on the Asiatic origin of these Indians. The distinct Gc subtype frequencies observed in our main ethnic groups provide an important new tool for anthropological analyses. Tf subtypes were studied among the Caingang only. The frequencies of Tf^C1 and Tf^C2 are similar to those obtained by other researchers in Hessen, Germany.     

The distribution of Gc subtypes among the Mongoloid populations

The polymorphism of Gc (group-specific components) has been investigated for a series of 3,160 individual samples from 11 Mongoloid populations in Asia and North and South America by isoelectric focusing on polyacrylamide gels. The samples fall into six Gc phenotypes which can be explained by the three common alleles, Gc^1F, Gc^1S, and Gc², together with several variant phenotypes explained as the heterozygotes for the three common alleles. The distribution of Gc^1F suballele appears to be considerably different from population to population among Mongoloids, ranging from 0.105 (Machiguenga Indans, Peru) to 0.609 (Kadazan, Borneo). A clear geographic cline from Southeast Asia into South America in Gc^1F allele was observed in the populations. In general, Gc^1F allele frequencies are lower in European populations and higher in African populations. The range of variability in the Gc^1F values observed among the Asiatic populations is between the Africans and the Europeans.     

Improving blast resistance of maintainer line DRR 9B by transferring broad spectrum resistance gene Pi2 by marker assisted selection in rice

Hybrid rice technology offers great promise to further enhance rice production and productivity for global food security. Improving hybrid rice parental lines is the first step in developing heterotic rice hybrids. To improve resistance against blast disease, a maintainer line DRR 9B was fortified with a major broad-spectrum blast resistance gene Pi2 through marker-assisted selection. The rice blast caused by Magnaporthe oryzae is a major disease and can cause severe yield losses upto 100%. The NILs of Samba Mahsuri namely BA-23-11-89-12-168 possessing Pi2 was utilized as a donor parent. The PCR-based molecular marker tightly linked to Pi2 gene was used for the foreground selection at BC₁F₁ generation. The molecular marker tightly linked to the major fertility restorer gene Rf4 was used for negative selection (i.e., selection of plants possessing non fertility restoring alleles) at BC₁F₁ generation to identify maintainer lines. The positive plants with Rf4 gene were added to the restorer pool for restorer line development. At each stage, MAS for Pi2 coupled with stringent phenotypic selection for agro-morphological and grain quality traits were exercised. At BC₁F₃ generation, one hundred families were screened against blast disease at uniform blast nursery (UBN) and selected resistant lines were advanced to next generations. In the BC₁F₅ generation plants were subjected to agro-morphological evaluation for yield and yield-contributing traits. The selected plants at BC₁F₅ generation were crossed with DRR 9A to assess the maintainer ability of blast resistance lines and for further CMS line conversion for hybrid rice breeding for developing blast resistance rice hybrids.     

Genetic polymorphism of the vitamin D binding protein (Gc protein) in pig plasma determined by agarose isoelectrofocusing

Genetic polymorphism of the pig plasma vitamin D binding protein Gc was demonstrated by agarose isoelectrofocusing followed by either autoradiography or immunofixation with specific human Gc antiserum. Three different types F, FS and S were observed. Family data supported the genetic theory that the Gc types are controlled by two autosomal codominant alleles Gc^F and Gc^s . Both alleles are present in Yorkshire and Duroc. In Danish Landrace and Hampshire only the Gc^F allele was observed.     

用环介导等温扩增技术快速检测粪便样本中的沙门菌

目的:建立快速检测粪便样本中的沙门菌的环介导等温扩增技术(LAMP),并着重在灵敏度和特异性方面对此方法进行评价。方法:利用LAMP针对沙门菌特定基因invA(靶基因)设计的6条特异引物,通过引物特异性识别特定基因invA上的8个独立区域来快速检测沙门菌;LAMP反应过程中会产生白色沉淀焦磷酸镁,故可以通过监测浊度来判定反应结果。结果:实时浊度仪监测反应结果表明,LAMP反应在60~65℃等温条件下50 min内完成;如果在反应前添加羟基萘酚兰,蓝色阳性结果很明显区别于紫色阴性结果;LAMP的最低检出限为6.97 pg/μL,PCR为69.7pg/μL,LAMP方法的检测灵敏度是PCR的10倍,且具有良好的特异性。结论:LAMP方法用于快速检测沙门菌,具有检测过程简单、实验装置简便、反应结果肉眼可辨、灵敏度高、特异性强的特点,对非沙门菌菌株的结果呈阴性,表明引物设计有很好的特异性。对粪便样本进行检测,发现具有同样的敏感性和特异性。这表明LAMP法是潜在的和有价值的在粪便样本中直接检测沙门菌的方法,具有快速、简便、低成本的特点。LAMP法适用于快速临床诊断。     

分子标记辅助选择聚合Xa23,Pi9和Bt基因

利用分子标记辅助选择将高抗水稻白叶枯病的Xa23基因、广谱高抗稻瘟病的Pi9基因、抗水稻螟虫和稻纵卷叶螟的Bt基因聚合到同一株系中,获得了三基因聚合的纯合株系。病、虫抗性鉴定结果显示：聚合了Xa23、Pi9和Bt基因的株系HB1471、HB1473能同时抗白叶枯病、稻瘟病和稻纵卷叶螟;与Xa23、Pi9基因的供体材料L10相比,对白叶枯病和稻瘟病的抗谱相同、抗性水平相当;对稻纵卷叶螟抗性与Bt基因的供体亲本MH63-Bt水平相当。Xa23、Pi9和Bt三基因纯合株系可以作为水稻育种的多抗供体材料。