Mitochondrial DNA deletion of the brain tissue of aged rats with learning and memory deficit

AIM and METHODS: The ratio of mitochondrial DNA (mtDNA) deletion was measured to find the relationship between mtDNA deletion and aged learning and memory deficit. The aged rats were divided into two groups, aged learning and memory deficit group and aged learning and memory normal group. The ratio of mtDNA deletion was measured by dilution polymerase chain reaction. RESULTS: There are deleted mtDNA (about 4834 bp) in the cerebral cortex, hippocampus and cerebellum of both young and aged rats. The ratios of deleted mtDNA were similar in the cerebral cortex,hippocampus and cerebellum of young rats (about 0.00018%). The ratio mtDNA of aged learning and memory normal rats had increased by five-fold in the cerebral cortex and hippocampus, or one-fold in the cerebellum over young rats. The ratio of aged learning and memory dificit rats had increased by one-fold in the cerebral cortex or 0.8-fold in the hippocampus or two-fold in the cerebellum over aged learning and memory normal rats.CONCLUSIONS: There was really the increase of mtDNA in aging rat brain. And this increase was double in amount in aged learning and memory deficit rats compared to the normal learning and memory aged rats. It is suggested that the mtDNA deletions in the brain regions associated with learning and memory may be contributed to the cellular and molecular mechanism of learning and memory deicit with aged rats.     

快速大量提取甜菜DNA的新方法

以甜菜幼嫩叶片为材料,通过冷冻真空干燥将材料变成干粉,利用CTAB法对DNA提取程序进行了研究,建立了一种简单、快速的甜菜DNA提取程序,既节省时间,又可获得高质量的DNA,可以满足SSR及SRAP扩增的需要。     

陆地棉SSR分子标记优化体系的建立

针对陆地棉富含酚类、多糖等次生产物的特点,建立了一种简便、快速的提取高纯度DNA的方法.同时建立了适合陆地棉SSR标记的优化体系和试验方案,并对所构建的QTL定位群体进行了SSRPCR检测,得到了清晰的多态性SSR标记,为SSR分子标记技术在棉花育种中的应用打下了基础.     

Polymorphism in Mitochondrial DNA of Six Fowl Breeds Revealed by Restriction Endonuclease

The patterns of cleavage of mtDNA by restriction endonucleases was analysed for six fowl breeds and of .Hyline White,Hyline Brown,ISA Brown,Hisex Brown,Lohmann White,Abor Acres and mtDNA polymorphisms were detected in the restriction patterns with the following eight enzymes,Ava Ⅰ,AvaⅡ,EcoRⅠ,HindⅢ,BamHI,PvuⅡ,PstⅠ,HincⅡ.The restriction cieavage patterns were identical among these breeds.Hyline White,Hyline Brown,ISA Brown,Hisex Brown,Lohmann White-A type.The patterns of Abor Acres were Btype.Based on their mtDNA restriction types,all the breeds were classified into two groups.Genetic distances among these groups were calculated in roder to define their phylogenetic relationships.The relationship among five egg line breeds is close,while Abor Acres(Broiler fow)is relatively far from them.The results suggest that the difference of mtDNA could result from the different origins.The polymorphic sites in mtDNA of Hyline White bas been located on a restriction map.     

Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers

True-to-type clonal fidelity is one of the most important pre-requisites in micropropagation of crop species. Genetic fidelity of in vitro raised 45 plants of gerbera (Gerbera jamesonii Bolus) derived from three different explants, viz., capitulum, leaf and shoot tips, was assessed by 32 ISSR markers, for their genetic stability. Out of 32 ISSR markers, 15 markers produced clear, distinct and scorable bands with an average of 5.47 bands per marker. The markers designed from AG motif amplified more number of bands. The markers anchored at 3′ ends produced high number of consistent bands than unanchored markers. Fifteen ISSR markers generated a total of 3773 bands, out of which 3770 were monomorphic among all the clones. The Jaccard's similarity coefficient revealed that out of 45 clones derived from different explants, 44 were grouped into a single large cluster alongwith the mother plant with a similarity coefficient value of 1.00, whereas one clone (C38) remained ungrouped. The clones derived from capitulum and shoot tip explants did not show any genetic variation, whereas, one of the leaf-derived clones exhibited some degree of variation.     

3种提取基因组DNA方法的比较分析

分别采用3种不同的方法提取基因组DNA，比较所提取的DNA的质量以及提取所需时间、费用，以期选择一种高效、简洁快速、价格合理的提取方法。结果发现这3种方法提取的质量都能达到相关分子生物学试验的要求，但在操作步骤、时间、价格等方面存在差异，具体比较结果可为分子生物学试验选择提取方法提供依据。     

苹果RAPD分析体系的建立

对富士苹果（Ｍａｌｕｓ　ｐｕｍｉｌａ　Ｍｉｌｌ．ｃｖ．Ｆｕｊｉ）ＲＡＰＤ（Ｒａｎｄｏｍ　Ａｍｐｌｉｆｉｅｄ　Ｐｏｌｙｍｏｒｐｈｉｃ　ＤＮＡ）分析体系的优化研究表明，ＲＡＰＤ反应体系中，ＤＮＡ、Ｔａｑ酶、引物和Ｍｇ２＋４种主要成分的最适用量分别为：２０ｎｇ、１．０ Ｕ、０．２ｕｍｏｌ·Ｌ－１和３．０ｕｍｏｌ·Ｌ－１。采用该优化体系，以 ＯＰＪ０３为引物，构建了我国及世界范围内苹果生产中重要的２８个品种的ＲＡＰＤ指纹图谱，分析了其遗传多样性，区分了供试的２８个苹果品种中的１５个，区分率达５３．６％。讨论了ＲＡＰＤ鉴定苹果品种的应用及其主要影响因素。     

Activation of repair pathways after neuronal DNA damage induced by bupivacaine

AIM To investigate the activation of related repair pathways after bupivacaine-induced neuronal DNA damage by cDNA gene screening. METHODS The bupivacaine-induced SH-SY5Y neuronal damage and DNA damage model was established. The technique of cDNA microplate array was used to screen the 21 important regulatory factors in the DNA damage repair pathway. Post-analysis of these differentially expressed repair genes for the repair pathway enrichment and distribution was performed. The data were analyzed by GraphPad Prism 6 statistical software to compare differences between groups. RESULTS The viability of SH-SY5Y cells treated with bupivacaine at different concentrations （detected by CCK-8 assay） showed that the IC₅₀ value of bupivacaine was 1.5 mmol/L. The comet assay related index （the comet tail） was increased （P<0.05）， the phosphorylation level of γH2AX protein was increased （P<0.05）， indicating that DNA damage in the SH-SY5Y cells was significantly aggravated after bupivacaine treatment. The results of cDNA microplate assay showed that compared withcontrol group， the differentially expressed genes after bupivacaine treatment were DNA-PKcs， PTEN， NTH1， RAD9， CSB， GADD45， XPD， XPC-HR23B and P53. The analysis showed that these repair genes were mainly concentrated in the following 3 repair mechanisms： base excision repair， nucleotide excision repair， and non-homologous reconstitution. CONCLUSION The repair genes differentially expressed after neuronal DNA damage caused by local anesthetics are mainly concentrated in the pathways of non-homologous end-joining， base excision repair and nucleotide excision repair.     

畜禽DNA指纹的研究与应用

综述了ＤＮＡ指纹技术的原理，方法，以及在个体（品种）鉴别，亲缘关系分析，群体遗传变异分析，连锁分析等方面的研究进展。     

表达H5亚型禽流感病毒HA基因的DNA疫苗免疫保护效力研究

 表达高致病力禽流感病毒分离株A/Goose/Guangdong/1/96(H5N1) [GD/1/96(H5N1)]HA基因的DNA疫苗质粒pCIHA5具有良好的免疫保护性,为了将其开发并应用于实际,对其免疫保护效力进行了进一步的研究。结果表明,用10、40、70、100和150g pCIHA5和油苗1次免疫后,其免疫保护率分别为12.5%(1/8)、58.3%(7/12)、72.7%(8/11)、50.0%(6/12)、66.7%(8/12)和100%(12/12),其中70和100g剂量组的病毒分离结果