牛胚胎在BRL/mTCM199和HECM-6/mTCM199培养体系中发育潜能的探讨

将受精卵置于下列两种培养体系中进行体外培养:(1)Buffalo rat liver细胞与改进的TCM199培养液共同培养系统(BRL/mTCM199);(2)半确定培养液两步培养系统[无血清的金黄地鼠培养液与改进的TCM199培养液(HECM-6/mTCM199]。结果表明:后者的囊胚率(50%)明显高于前者的囊胚率(41%,P<0.01)。表明牛胚胎早期分裂可在无蛋白培养条件下进行,并且在此培养条件下有良好的后续发育能力。这可能与两步培养体系中葡萄糖和氨基酸浓度不同有关。

基于部分线粒体细胞色素氧化酶亚基Ⅰ(mtCOI)DNA 序列的6种蝙蝠(翼手目:蝙蝠科)的分子系统进化关系

对贵州5种蝙蝠科蝙蝠的部分线粒体细胞色素氧化酶亚基Ⅰ DNA序列进行了测定,并结合从Genbank获得的爪哇伏翼的相应序列,以Pteropus dasymallus,P.scapulatus,Rousettus aegyptiacus为外群,运用贝叶斯法(Bayesian),最大似然法(Maximum Likelihood,ML)分析了这6种蝙蝠科蝙蝠的分子系统进化关系.结果表明:在贝叶斯,ML树中,这6种蝙蝠科的蝙蝠可分为3个分支:亚洲长翼蝠是第1个独立出来的分支;白腹管鼻蝠是继亚洲长翼蝠之后第2个分离出来的分支;第3个分支又分为两支,一支由大鼠耳蝠和小鼠耳蝠组成,另一支由南蝠和爪哇伏翼组成,支持将这4种蝙蝠同归于蝙蝠亚科的结论,从一定程度上否定了鼠耳蝠属和管鼻蝠亚科之间的姐妹类群关系,也不支持将鼠耳属提升为亚科.

6种常用渔药对厚颌鲂鱼苗的急性毒性试验

在水温29~30℃条件下,用硫酸铜与硫酸亚铁合剂(比例5:2)、高锰酸钾、晶体敌百虫、强氯精、溴氯菊脂、甲醛共6种药物对厚颌鲂(Megalobrama pellegrini)鱼苗进行急性毒性试验。试验结果显示,6种药物48h的半致死浓度分别为5.78、6.50、4.90、2.55、0.042、120mg/L,安全浓度分别为0.445、0.528、0.268、0.190、0.003、7.500mg/L,厚颌鲂鱼苗对药物敏感性比一般鱼类高,实际用药时应根据具体情况确定用药时间和用药量,溴氯菊脂不宜作为治疗药物

鲤鱼3个亚种线粒体DNA ND5/6区段的PCR-RFLP分析及亚种间分子标记的确立

从鲤鱼3个亚种(Cyprinus carpio carpio,Cyprinus carpio haematopterus和Cyprinus carpiorubrofuscus)中选出具代表性的品系共10个,运用PCR方法,扩增出2.4 kb的mtDNA ND5/6区段.共用9种限制性内切酶进行酶切分析,结果表明,每个亚种拥有一种单倍型,有4种限制性内切酶(Dde I,Hae III,Taq I和Mbo I)酶切后产生了能将3个亚种区分开来的诊断性限制性酶切位点.可利用其作为鉴定3个亚种的遗传标记和遗传育种

6种累枝虫(Epistylis)rRNA基因18S-ITS1序列及其分子系统发育关系

建立了由采自自然界中的样品、不经实验室培养而直接用于大核DNA提取和PCR反应的原位（in situ）方法;在此基础上,测定并比较了6种累枝虫（Epistylis wenrichi,E.urceolata,E.chrysemydis,E.plicatilis,E.hentscheli,E.galea）的18S-ITS1序列,结果显示:E.wenrichi,E.urceolata,E.chrysemydis,E.plicatilis和E.hentscheli间18S和ITS1区序列的碱基相似性很高,而它们与

鲤科鱼类6个正反交人工异源三倍体胚胎的染色体变化及其发育命运

采用静水压休克保留第二极体的方法,在鲤(♀)×鲢(?)、鲫(♀)×鲢(?)、白鲫(♀)×鲢(?)和鲢(♀)×鲤(?)、鲢(♀)×鲫(?)、鲢(♀)×白鲫(?)6个正反交组合中都诱导出了异源三倍体,但只有正交鲤(♀)×鲢(?)、鲫(♀)×鲢(?)和白鲫(♀)×鲢(?)3个处理组中整倍性的异源三倍体胚胎才有可能正常发育,孵化出苗;而反交鲢(♀)×鲤(?)、鲢(♀)×鲫(?)和鲢(♀)×白鲫(?)3个处理组中的异源三倍体胚胎在发育过程中不断发生染色体排除和丢失,形成非整倍体而死亡,只有少数雌核发育二倍体鲢才能孵

果糖1,6-二磷酸酯酶的代谢调控

<正> 果糖1,6-二磷酸酯酶(FBPase 1)在生物体内糖异生代谢途径中起着重要的作用。当二价金属离子存在时,它催化果糖1,6-二磷酸(Fru-1,6-P_2)水解生成果糖6-磷酸(Fru-6-P)和无机磷。它是一个由4个相同亚单位构成的蛋白质,其亚单位分子量约为35 000～40 000。FBPase 1可被蛋白水解酶降解修饰,使其最适pH从7.5变为9.2。该酶受AMP及果糖2,6-二磷酸(Fru-2,6-P_2)

Waterborne exposure to fluorotelomer alcohol 6:2 FTOH alters plasma sex hormone and gene transcription in the hypothalamic-pituitary-gonadal (HPG) axis of zebrafish

Fluorotelomer alcohols (FTOHs) have shown estrogenic activity in vitro and in vivo, but the mechanism of this activity is not known. In this study, 18-week-old zebrafish (Danio rerio) were exposed to 0, 0.03, 0.3 and 3.0 mg/l 1H, 1H, 2H, 2H-perfluorooctan-1-ol (6:2 ETCH) for 7 days, and the effects on plasma sex hormone levels were measured followed by use of real-time PCR to examine selected gene expression in hypothalamic-pituitary-gonadal (HPG) axis and liver. Exposure to 6:2 FTOH significantly increased plasma estradiol (E2) and testosterone (T) levels in both males and females. Furthermore, the ratio of T/E2 was reduced in females while increased in males. In females, the increase of E2 was accompanied by up-regulated hepatic estrogenic receptor alpha (ER alpha) and vitellogenin (VTG1 and VTG3) expression. In males, the elevation of the T level is consistent with the up-regulation of cytochrome P450 c17 alpha-hydroxylase, 17, 20-lase (CYP17) and the down-regulation of cytochrome P450 aromatase A (CYP19A). The present study demonstrated that waterborne exposure to 6:2 FTOH alter plasma sex hormone levels and the ratio of T/E2, as well as the transcriptional profiles of some genes in the HPG axis and liver. The results suggested that FTOHs may disturb fish reproduction through endocrine disrupted activity. (C) 2009 Elsevier B.V. All rights reserved.

RNAi suppression of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) mediated differentially expressed genes involved in Toll-like receptor signaling pathway and caused increased susceptibility to Flavobacterium columnare

In Drosophila, Toll signaling cascade, which resembles the mammalian Toll-like receptor (TLR)/IL-1R signaling pathways and regulates the expression of anti-microbial peptide genes, mainly relies on peptidoglycan recognition proteins (PGRPs) for the detection of bacterial pathogens. To explore the effect of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) on Toll-like receptor signaling pathway, RNA interference (siRNA) and real time quantitative PCR (RQ-PCR) methods were used to identify differentially expressed genes regulated by zfPGRP6. The target genes included TLR2, TLR3, TLR5, TLR7, TLR8, IL1R, Sterile-alpha and Armadillo motif containing protein (SARM), myeloid differentiation factor 88 (MyD88) and nuclear factor (NF)-kappa B2 (p100/p52). The results of RQ-PCR showed that RNAi-mediated Suppression of zfPGRP6 significantly down-regulated the expression of TLR2, TLR5, IL1R, SARM, MyD88 and p100/p52. The expression of beta-defensin-1 was also down-regulated in those embryos silenced by zfPGRP6. In challenge experiments to determine the anti-bacterial response to Gram-negative bacteria, RNAi knock-down of zfPGRP6 markedly increased susceptibility to Flavobacterium columnare. (C) 2008 Elsevier B.V. All rights reserved.

PCR-RFLP analysis of mitochondrial DNA ND5/6 region among 3 subspecies of common carp (Cyprinus carpio L.) and its application to genetic discrimination of subspecies

Mitochondrial DNA ND5/6 region was studied by PCR-RFLP analysis among ten representative strains belonging to three subspecies (Cyprinus carpio carpio, Cyprinus carpio haematopterus and Cyprinus carpio rubrofuscus) of common carp (Cyprinus carpio L.). A total of 2.4 kb fragment was amplified and subjected to restriction endonuclease analysis with nine restriction endonucleases subsequently. The results indicated that each subspecies owned one hyplotype and four restriction enzymes (Dde I, HaeIII, Taq I and Mbo I) produced diagnostic restriction sites which could be used for discriminating the three subspecies and as molecular genetic markers for assistant selective breeding of common carp.

Isolation by phage display and characterization of a single-chain antibody specific for O-6-methyldeoxyguanosine

New approaches of making single chain Fv antibodies against O-6-methyl-2'-deoxyguanosine (O(6)MdG) have been demonstrated by using the phage antibody display system. Using O(6)MdG as an antigen, 21 positive clones were identified by ELISA from this library, one of which, designated H3, specifically binds to O(6)MdG with high affinity. The H3 scFv antibody has an affinity constant (K-aff) of 5.94 x 10(11)(mol/L)(-1). H3 scFv has been successfully used to detect O-6 MdG in DNA hydrolyses from yeast or E. coli cells treated with a DNA methylating agent. To our knowledge, this is the first report of the selection of a specific scFv against DNA adducts. The results demonstrate the potential applications of the phage display technology for the detection of DNA lesions caused by mutagens and carcinogens.