Genetic study of Kelp “901” strain

Based on DNA extraction and optimization of random amplified reaction (RAPD) to the gametophytes and sporophytes of Kelp “901” strain, genetic study on variation was conducted to its parents and offsprings of F6, F7,Fs, and F9 generation. RAPD results have shown that among 30 selected primers for gametophytes, 297 loci ranging from 200 to 3 000 bp were obtained in the average of 9,9 loci for each primer, This indicated a high polymorphic rate with RAPD detection. UPGMA (unweighted pair-group method arithmetic average) analysis showed that each male and female gametophyte of a generation could be clustered into one pair separately. The genetic distances of the Kelp 901 generation were 0.321 2-0.476 7, and the maximum was between F7 and Fs (0.476 7). Identity analysis showed that F6 generation was more close to the female parent (0.659 3), and F7 generation was more close to the male parent (0.5788). To the sporophytes study in 24 selected primers for RAPD amplification, 191 loci ranging from 230-2 800 bp were obtained, in the average to each primer of 8.0 loci. The heterozygosity to six populations were male parent (0.223 9),female parent (0.107 2), F6 (0.216 4), F7 (0.228 6), F8 (0.229 6) and F9 (0.317 2). The nearest genetic distance was 0.083 5 (Fs, F9). Total heterozygosity (HT) ofF6, F7, F8 and F9 generations was 0.318 6, the average heterozygosity (Hs) for F6,F7, F8 and F9 generations was 0.248 0, and deduced coefficient of population differentiation (Gst) was 22.2%. Six sequence characterized amplified regions (SCAR) were preliminary screened through RAPD analysis. It needed to be verified in detail as they are significant for molecular marker assistance in breeding and selecting Laminaria.     

Effects of heavy metal ions (Cu2+, Pb2+ and Cd2+) on DNA damage of the gills, hemocytes and hepatopancreas of marine crab, Charybdis japonica

There are rising concerns about the hazardous effects of heavy metals on the environment. In this study, comet assay and DNA alkaline unwinding assay were conducted on the tissues (gills, hepatopancreas, and hemocytes) of Charybdis japonica in order to illustrate genotoxicity of three heavy metal ions (Cu2+, Pb2+, and Cd2+) on the marine crabs C. japonica. The crabs were exposed to Cu2+ (10, 50, and 100 ?g.L?1), Pb2+ (50, 250, and 500 ?g L?1) and Cd2+ (5, 25, and 50 ?g L?1), and the tissues were sampled at days 0.5, 1, 3, 6, 9, and 15. DNA alkaline unwinding assay was used for testing the DNA single strand break in gills and hepatopancreas and comet assay was employed for testing the DNA damage in hemocytes. The results showed that the DNA damage (F-value) of gills in the crabs exposed to the three heavy metals was decreased gradually during the exposure periods and there was a dose-time response relationship in certain time, suggesting that the levels of DNA single strand break in all the experimental groups increased significantly compared to the controls. Changes of F-value in hepatopancreas of the crabs exposed to the three heavy metals were similar to those in gills except that the peak values were found in the 500 ?g L?1 Pb2+ treatment group at day 3 and the 50 ?g L?1 Cd2+ treatment group at day 9. The ranks of DNA damage in gills and hepatopancreas induced by the three heavy metal ions (50 ?g L?1, day 15) were Cd2+ >Pb2+ >Cu2+ and Pb2+ >Cu2+ >Cd2+. The levels of DNA damage in gills were higher than those in hepatopancreas in the same experimental group. It can be concluded that indices of DNA damage can be used as the potential biomarkers of heavy metal pollution in marine environment.     

双链DNA片段的碱基空间模式BSP——基因组调控区的一种可能的信息编码方式

生物体可看作是一种特殊的信息处理系统 ,论文的第二章从这种观点出发讨论了生物细胞内最大的信息源基因组DNA序列的信息编码问题。首先探讨了基因组蛋白质编码区的信息编码问题 ;简要回顾转录水平的基因表达调控后 ,讨论了基因组基因表达调控区的信息编码问题。第三章介绍了我们利用internet上可获取的开放的生物信息学资源 ,对基因组DNA调控区的一种可能的编码方式所进行的理论研究。这种可能的编码方式我们称之为双链DNA上的碱基空间模式BSP (BaseSpatialPattern)。首先 ,我们发现了由三联体BSP介导的一种 4与 2 0的关系 ,称为 4 -BSP -2 0关系。并推测 4 -BSP -2 0关系也许是某些DNA—蛋白质相互作用中的一种识别密码 ;第二 ,研究真核细胞转录调控中的DNA—蛋白质相互作用的实验数据表明 :某些DNA结合蛋白质对双螺旋核酸的专一性识别可能与BSP有关。并找到了 7组由其他生物学家公开发表的实验数据支持这一猜测 ;第三 ,研究了BSP的形式化描述问题 ,找到了一种基于等价类的BSP描述方法 ,并开发了相应的计算机处理算法。该描述方法简明且便于用计算机进行处理 ,更重要的是它提供了一种研究途径 ,可把BSP和 4 -BSP -2 0关系推广到单链DNA (ssDNA)和ssDNA -蛋白质相互作用的研究中 ,从而可进一步推广到RN     

应用巢式逆转录聚合酶链反应检测鱼类病毒性出血性败血症病毒(VHSV)

参照鱼类病毒性出血性败血症病毒序列,根据PCR引物设计的原则,设计巢式PCR引物,采用巢式逆转录聚合酶链反应(RT-PCR)方法,对如何快速检测鱼类病毒性出血性败血症病的方法进行了较为系统的研究,并对RT-PCR的反应条件进行了优化.结果表明,巢式RT-PCR扩增获得279bp的特异性片断,阴性对照无扩增条带.巢式RT-PCR扩增出的特异性片段经测序分析,结果证实与报道的序列完全一致.该方法最低可检测出0.1pg的鱼类病毒性出血性败血症病毒RNA.初步建立了VHSV的巢式RT-PCR检测方法,该方法灵敏、特异,可为VHSV的检测提供一个快速、有效的手段.     

5个湖泊河川沙塘鳢(Odontobutis potamophila)种群线粒体细胞色素b基因的遗传变异分析

通过对5个湖泊的河川沙塘鳢种群的线粒体DNA细胞色素b基因进行PCR扩增、测序,获得1141 bp的序列全长.序列分析显示,cyt b基因序列中A+T含量(55.8%)略高于G+C含量(44.2%),共检测到806个多态位点,115个样本得到87个单倍型,平均单倍型多样性为0.969±0.012,核苷酸多样性为0.20081±0.00742,遗传多样性表现高度多样性.太湖种群与大纵湖种群间的遗传距离最近,为0.137,巢湖种群和大纵湖种群之间遗传距离最远,为0.424.分子方差分析表明,群体间遗传分化系数Fst为0.531,变异来自群体内及群体间.cyt b基因序列构建的UPGMA系统进化树显示,5个种群分化成不同的分支系谱,种群间存在的基因交流较少.     

无机离子（Ni,Ge）和化剂对DNATm值的影响

在实验条件下，选择几种有代表性的无机离子，并保持一定的浓度和ＰＨ值，研究了各无机离子烷化剂对ＤＮＡＴｍ值的影响。结果表明，有益元素与有害元素对ＤＮＡＴｍ值高低的影响明显不同。     

转“全鱼”溶菌酶基因大菱鲆的研究

用大西洋条鳕(Macrozoarces americanus)抗冻蛋白启动子opAFP promoter和牙鲆(Paralichthys olivaceus)c型溶菌酶基因构建“全鱼”溶菌酶基因元件opAFP—ly，首次应用电脉冲精子介导法将该基因片段导入到大菱鲆(Scophthalmus maximus)受精卵内，获得原代转基因大菱鲆。先后采用单因子方法和正交实验方法，重复实验确定了最适导入条件：脉冲电压为400V／cm，脉冲次数为5，脉冲宽度为25ms，外源DNA浓度为50mg/L。利用最适导入条件，实现基因元件大规模转移。采用PCR技术对1月龄的原代转基因大菱鲆仔鱼的染色体DNA进行分析，结果表明外源基因的整合率可达28％。     

大口鳒线粒体DNA控制区结构和鲽形目鱼类的系统进化初步研究

测定大口鳒Psettodes erumei线粒体控制区全序列,并与其他7种鲽形目鱼类控制区结构进行比较分析.结果表明,大口鳒线粒体控制区全长大约1601bp,5'端和3'端分别存在56bp和8bp的串联重复序列.除此之外,其控制区结构和其他种类相似,分为终止相关序列区(TAS)、中央保守区(CSB-D、CSB-E、CSB-F)、保守序列区(CSB-1、CSB-2、CSB-3),同时在CSB-D后还识别出鲽形目保守的Poly-T结构.比较了8种鲽形目鱼类控制区全序列和去除重复序列后控制区序列的碱基组成变化,结果显示前者的种间差异明显大于后者,变异系数约是后者的两倍.这种差异主要是由于重复序列的异质性如长度及其拷贝数不同、碱基组成不同所造成的.基于控制区部分序列对鲽形目12种鱼类进行系统分析,结果显示,在鲽形目内部大口鳒分类地位最原始;鲽亚目先与鳎亚目聚在一起,后与鳒亚目聚在一起,这个结果与基于形态特征的系统关系相符.     

Application of the first internal transcribed spacer(ITS-1) of ribosomal DNA as a molecular marker to population analysis in farrer''s scallop Chlamys farreri

Sequence variation of the first internal transcribed spacer of ribosomal DNA （ ITS - 1 ） was examined and its application to the study of genetic variation was explored in four populations of farter＇ s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA （analysis of molecular variance） showed that the majority （96.26%） of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 （ = 0.042）, indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.     

北京西北城区一次重污染过程中PM10的氧化损伤能力研究

为评估大气重污染期间北京市可吸入颗粒物(PM_(10))的毒性,采集2016年3月份一次大气重污染过程中北京市大气PM_(10)样品,应用质粒DNA损伤评价法来研究其氧化损伤能力。结果表明,雾霾期间PM_(10)对DNA的损伤率高于雾霾逐渐消退时期,远高于雾霾前期清洁天,颗粒物对DNA损伤率随剂量的增加而增加。雾霾前后PM_(10)水溶样品的平均TD_(20)值表现为雾霾前清洁天(788.01μg/mL)雾霾消退后期(470.40μg/mL)雾霾期间(55.78μg/mL),说明氧化能力雾霾期间雾霾消退后期雾霾前清洁天。另外通过雾霾前后数据对比得到,暴露毒性指数TI大小顺序为雾霾期间(13 245.06)雾霾消退后期(1 658.87)雾霾前清洁天(254.08),说明雾霾期间PM_(10)对人体危害更大。