共查询到19条相似文献,搜索用时 62 毫秒
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应用红外荧光和加尾引物法进行向日葵SSR遗传分析中的多聚PCR和多聚凝胶电泳的优化 总被引:2,自引:0,他引:2
在向日葵(Helianthus annuus L.)自交系SSR遗传分析中,为了提高SSR荧光分析效率、简化分析程序和降低研究费用,我们进行了多聚PCR和多聚凝胶电泳两项技术的优化研究。结果表明,优化多聚PCR和多聚凝胶电泳的影响因子(如逐步降低的退火温度的循环数、各个多聚位点引物浓度的平衡、PCR缓冲液浓度以及Taq DNA多聚酶的浓度等)可以收到更好的实验效果。基于以上的优化研究,系统地提出了一套优化的加尾引物法的策略。同时,提出了在多聚PCR和多聚凝胶电泳中常常遇到的技术难题的有效防止和解决的方法。 相似文献
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多聚唾液酸与多聚唾液酸转移酶 总被引:3,自引:0,他引:3
多聚唾液酸(PSA)是一种在神经细胞黏附分子(neural cell adhesion molecule,NCAM)上表达的唾液酸聚合物,在神经发育过程中起重要作用.PSA的聚合程度会影响PSA-NCAM的功能.多聚唾液酸酶主要用于合成PSA-NCAM,两种高度同源的多聚唾液酸转移酶ST8SiaⅡ和ST8SiaⅣ都属于唾液酸转移酶家族.多聚唾液酸转移酶中NCAM的识别域和多聚唾液酸化域是截然不同的,且一些异构酶在NCAM多聚唾液酸化中起明显的负作用.多聚唾液酸酶与很多疾病都有关系,以多聚唾液酸转移酶为标靶设计的药物也将成为神经系统及肿瘤治疗的新型药物. 相似文献
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羽叶金合欢的DNA提取和SSR引物筛选 总被引:2,自引:0,他引:2
利用SSR分子标记对羽叶金合欢( Acacia pennata )的野生种和栽培种进行了分析,利用改良的CTAB方法优化了其总DNA的提取方法,并优化了PCR扩增条件.从已有的金合欢属植物的82对SSR引物中筛选出多态性高,稳定性好的12对引物作为羽叶金合欢的SSR分析引物,为进一步对其进行遗传多样性研究提供了依据. 相似文献
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目的:比较L型多聚赖氨酸(L-PL)和D型多聚赖氨酸(D-PL)浸泡的玻片培养小鼠大脑皮层神经元细胞对其分化的影响。方法:取昆明白小鼠的大脑皮层细胞,分别用L-PL和D-PL进行培养,统计和比较细胞的分化比率、突起数量以及突起生长长度。结果:L-PL和D-PL培养的神经细胞第1、2天的分化率比较差异具有统计学意义(P0.05),而突起数和突起长度比较差异无统计学意义(P0.05)。当L-PL、D-PL的浓度分别为20μL/m L、5μL/mL时,其对1、2天细胞的分化产生显著影响,并且D型多聚赖氨酸培养的细胞分化率高于L型。500、100、20μg/m L的L-PL中,100μg/ml L-PL培养细胞的分化率最高,而125、25、5μg/m L的D型多聚赖氨酸培养细胞分化率比较差异虽然无统计学意义,但25μg/ml D-PL培养神经细胞的分化率总体趋势高于其他浓度。D-PL所形成的粒径大小与L-PL不同,且D-PL的总体趋势稍大于L-PL。结论:L型多聚赖氨酸和D型多聚赖氨酸会在神经细胞生长早期对分化率产生影响,但不影响突起数和突起长度。 相似文献
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亚硝酸钠和多聚磷酸钠对日本沼虾的毒性研究 总被引:8,自引:0,他引:8
针对目前危害虾类养殖的水污染中比较严重的亚硝酸盐和多聚磷酸盐污染,主要研究亚硝酸钠和多聚磷酸钠对日本沼虾的毒性影响。通过实验得出:亚硝酸钠对日本沼虾24、48、96h的半致死浓度分别为46、26、13.33mg/L,多聚磷酸钠对日本沼虾24、48、96h的半致死浓度分别为l233.33、1180、1080mg/L。实验表明,活性氧是随着亚硝酸钠和多聚磷酸钠的浓度增加而增加,而超氧化物歧化酶的酶活力则是随着亚硝酸钠和多聚磷酸钠的浓度增加先稍微增加,后又下降,这表明虾的免疫系统对低浓度的毒物有一定的抵抗能力,但随着毒物浓度增大,虾的免疫系统会受到抑制,虾的免疫功能下降,抗病力降低,导致染病死亡。 相似文献
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利用SSR 分子标记对羽叶金合欢( Acacia pennata) 的野生种和栽培种进行了分析, 利用改良的CTAB方法优化了其总DNA 的提取方法, 并优化了PCR 扩增条件。从已有的金合欢属植物的82 对SSR 引物中筛选出多态性高, 稳定性好的12 对引物作为羽叶金合欢的SSR 分析引物, 为进一步对其进行遗传多样性研究提供了依据。 相似文献
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多聚磷酸盐及其代谢酶的研究进展 总被引:1,自引:0,他引:1
多聚磷酸盐(polyP)是由几个到几百个无机磷酸盐单体通过高能磷酸键聚合而成的线性多聚体,广泛分布于自然界和生物体.本文总结了polyP在生物体中的重要功能,包括基因表达和调控、DNA的摄取、微生物的运动性、对胁迫和饥饿的应答、病原菌的毒性以及对细胞凋亡、血液凝固、细胞钙化、线粒体功能的调节,需要polyP的酶有内切酶、葡萄糖激酶、NAD激酶和AMP磷酸转移酶等.本文对调控polyP的多聚磷酸盐激酶(polyphosphate kinase,ppk)和外切聚磷酸酶(exopolyphosphatase,PPX )的生化性质和结构也进行了总结.同时,结合我们的研究工作,重点分析了结核分枝杆菌中PPX的同源蛋白和可能的生物化学活性. 相似文献
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猪微卫星标记多重PCR扩增组合 总被引:1,自引:3,他引:1
采用多重PCR的方法,以快速扩增微卫星标记和节约试剂为目标,对其反应条件进行优化后,获得了46个扩增效果理想的微卫星标记多重PCR组合,其中30个为二重PCR,16个为三重PCR。实验结果表明,这些多重PCP反应的引物浓度为0.06~0.3μmol/L, Mg2+的浓度变化范围为1.5~3.0mmol/L,采用的PCR缓冲液的倍数为1.0、1.2、1.4或1.6,每个PCR反应聚合酶的用量在0.2和0.4U之间,退火温度及反应循环数分别为52~60℃ 和32~50℃。所有多重PCR进一步合并为17个可在ABI 337测序仪上进行电泳的组合。
Abstract:In order to rapidly amplify pig microsatellite markers and save materials,multiplex PCR was used and its reaction condition was optimized.Forty-six combinations of multiplex PCR with good effects were obtained.Thirty of them are duplex-PCRs,sixteen are triplex-PCRs.The results of multiplexes showed that the concentration of primers varied among 0.06~0.3μmol/L,the Mg2+ concentration among 1.5~3.0 mmol/L;0.2~0.4 U of Taq polymerase and 1.0-,1.2-,1.4-,1.6-fold buffer were used,the annealing temperature and the cycle number varied among 52~60℃ and 32~50℃,respectively.All multiplexes were further combined into 17 sets for the electrophoresis on ABI 377 sequencer. 相似文献
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LIU Qiu-Ling LÜ De-Jian ZHU Jia-Zhen LU Hui-Ling LUO Yan-Min FANG Qun 《Acta Genetica Sinica》2006,33(12):1053-1059
This study was carried out to evaluate the value of three X-STR loci (DXS6803, DXS981and DXS6809) in forensic application and thereby investigate their polymorphism. The primer for each locus was labeled with fluorochrome 6-FAM. A fluorescent multiplex PCR for simultaneously amplifying three X-STR loci was set up. The PCR products that were obtained were analyzed using capillary electrophoresis and ABI PRISM 3100 Genetic Analyzer, with GENESCAN Analysis Software. When 340 male and 195 female individuals of Han population in China were tested, 13, 12, and 11 alleles were observed for DXS6803, DXS981 and DXS6809, respectively. One hundred and eighty three haplotypes were detected in the male individuals. The haplotype diversity reached 0.9926. The results show that the three loci of the multiplex system provide significant information on polymorphism for forensic identification and paternity testing, particularly for complicated paternity deficient cases. 相似文献
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Schuenck RP Pereira EM Iorio NL Dos Santos KR 《FEMS immunology and medical microbiology》2008,52(3):431-435
Staphylococcus haemolyticus is the most frequently coagulase-negative Staphylococcus species associated with antimicrobial resistance isolated from nosocomial infections. We developed an accurate and simple multiplex PCR assay to identify methicillin-resistant S. haemolyticus (MRSH) isolates. We designed species-specific primers of the mvaA gene that encodes a 3-hydroxy-3-methylglutaryl coenzyme A involved in the mevalonate pathway of the microorganism. Simultaneously, mecA gene primers of methicillin resistance were also used. The PCR assay was established using 16 strains of different reference Staphylococcus species and validated with a collection of 147 clinical staphylococcal isolates that were also phenotypically characterized. Reliable results for the detection of MRSH isolates were obtained for 100% of the strains evaluated, showing that this PCR assay can be used for the routine microbiology laboratories. This is the first report using species-specific multiplex PCR to detect a single segment of S. haemolyticus associated with a segment of mecA gene. 相似文献
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4个Y-STR基因座银染复合扩增 总被引:10,自引:2,他引:8
建立一个复合扩增体系,同时扩增DYS391、GATA-A4、GATA-A10和GATA-H4 4个Y染色体特异性的STR基因座,并用聚丙烯酰胺凝胶电泳和银染显色技术进行基因分型。采用该复合扩增系统调查广东汉族311名无关男性个体的单倍型频率,这4个基因座分别检出5、7、6和5个等位基因,其基因多样性分别为0.4623、0.6972、0.7173、0.6015。共检出98种单倍型,单倍型基因多样性为0.9755。该复合扩增体系在建立Y染色体STR数据库,研究群体遗传和进行法医学鉴定有重要意义。
Abstract:A multiplex PCR system has been developed to amplify 4 Y-chromosome specific short tandem repeats(STR),DYS391,GATA-A4,GATA-A10 and GATA-H4,simultaneously.PCR products were separated by polyacrylamide gels electrophoresis followed by silver stain.When 311 unrelated males from the Han population in Guangdong were detected by the multiplex system,DYS391,GATA-A4,GATA-A10 and GATA-H4 showed 5,7,6 and 5 alleles respectively.Total 98 haplotypes could be identified.Gene diversity value for the 4 STR was 0.4623,0.6972,0.7173 and 0.6015 respectively.The gene diversity value for the haplotypes of the 4 Y-STR reached 0.9755.The four Y-STR multiplex system will be very powerful for establishing Y-STR database,exploring human origin,paternity testing and personal identification. 相似文献
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Following the recent emergence of malaria in South Korea, vector control has been an important task. For this, vector identification is very important. Earlier, two PCR-based assays have been described. But, poor species resolution and their ability to include only 4-5 species limit their use. Thus, it has now become important to revise the assay identifying these members. In this study, a new assay based on internal transcribed spacer 2 and 28S of ribosomal DNA has been described. The assay successfully identified all the Korean malaria vector mosquitoes. Therefore, it is an indispensable tool to study ecology, abundance and biology of these species. 相似文献