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151.
【目的】研究禽源奇异变形杆菌携带ampC基因的分型和blaCMY-2阳性接合质粒pC12的序列结构,为防控多重耐药禽源奇异变形杆菌的传播提供理论基础。【方法】利用头孢西丁三维试验和PCR方法对21株奇异变形杆菌进行AmpC酶的检测和基因分型研究;对blaCMY-2阳性菌株进行脉冲场凝胶电泳(PFGE)分型和接合试验;利用高通量测序技术获得pC12的核苷酸序列并进行比较分析。【结果】头孢西丁三维试验表明21株禽源奇异变形杆菌中有6株产AmpC酶。6株产AmpC酶奇异变形杆菌对氨苄西林、头孢西丁、多西环素、氟苯尼考、粘菌素全部耐药,而对头孢他啶、阿米卡星全部敏感。耐药基因PCR扩增和测序结果表明,6株菌均携带blaCMY-2,检出率为28.6%。接合试验表明,1株奇异变形杆菌C12接合成功并获得接合子,其余5株接合不成功。PFGE分型结果表明,酶切图谱分为3个型别,blaCMY-2在禽源变形杆菌中存在垂直和水平传播。测序结果表明菌株C12含有一个1b型IncC质粒,其全长161 319 bp,GC含量为52.45%,预测有161个开放阅读框,提交NCBI并获得序列号MT320534。该质粒包含3个耐药区:第一个抗生素耐药区(antibiotic resistance islands,ARI-B)携带floR、tet(A)、strA、strB和sul2;第二个耐药区是一个典型的ISEcp1-blaCMY-2-blc-sugE结构,其中的ISEcp1被一个插入的IS10R截断;第三个耐药区(ARI-A)是一个杂合的Tn1696tnp-pDUmer转座子,包含一个1类整合子基因盒(aac(6')-Ib-cr|arr3|dfrA27|aadA16)和汞抗性基因簇merEDBAPTR,其插入到质粒骨架产生两个重复序列TTGTA,该耐药区也是1型IncC耐药区变化最大的区域。【结论】6株产AmpC酶禽源奇异变形杆菌均携带blaCMY-2,其酶切图谱分为3个PFGE型别,其中一株菌携带了一个流行的blaCMY-2阳性1型IncC可接合质粒。广宿主的IncC质粒是blaCMY-2、tet(A)、floR等多个耐药基因及整合子的重要载体之一,该质粒在动物源奇异变形杆菌的传播扩散进一步增加了治疗该菌感染的难度,应引起足够的重视。 相似文献
152.
卵泡抑素(follistatin,Fst)蛋白具有拮抗TGF-β超家族许多成员的功能,通过直接的蛋白结合可抑制肌肉抑制素(myostatin)的活性,从而恢复肌肉的生长,表现出增肌效应。参照鲤高通量转录组测序数据的拼接结果,筛选出鲤卵泡抑素Fst1基因的编码框(ORF)序列(全长1 260 bp,编码320个氨基酸),构建了包含金鱼Tgf2转座子左臂(220 bp)、右臂(185 bp)、斑马鱼Mylz2启动子和鲤Fst1基因ORF的供体质粒pTgf2-Mylz2-ccfst1。通过显微注射方法,并在体外合成的Tgf2转座酶5'加帽mRNA的介导下,获得了一批转鲤Fst1基因的转基因鲤。经PCR检测,转基因鲤Fst1外源基因的整合率平均为44.7%,对其中4尾转基因阳性鲤的扩增产物进行回收、克隆和测序验证,结果表明阳性鲤中均含有转基因目的片段。说明Tgf2转座子转基因系统可在鲤中实现较高的转座效率,为进一步研究卵泡抑素在鲤肌肉发育中的功能奠定了基础。 相似文献
153.
Pascal Bach Laurent Dagorn Arnaud Bertrand Erwan Josse Christophe Misselis 《Fisheries Research》2003,60(2-3):281-292
This study compares detailed, nearly continuous, observations on bigeye tuna, Thunnus obesus equipped with electronic tags, with discrete observations on a larger number of individuals from fishing experiments in order to validate the use of instrumented longlines to study the vertical distribution of fish. We show that the depth distributions obtained from the two different observation techniques regarding different environmental variables (temperature, dissolved oxygen (DO), prey distribution) are similar. Bigeye tuna do not seem to be attracted by baits in the vertical dimension (no modification of their vertical distribution by the fishing gear), which allows the use of instrumented longlines to study the vertical behaviour of pelagic species. This technique, when used with appropriate deployment strategy, could therefore represent an alternative to electronic tags (acoustic or archival tags) when there is a need to determine the vertical distribution of fish species by size or sex, in different environments for the study of fishery interactions. 相似文献
154.
Nicolás Astorga Juan Manuel Afonso María Jesús Zamorano Daniel Montero Víctor Oliva Hipólito Fernández & María Soledad Izquierdo 《Aquaculture Research》2005,36(8):733-738
The use of the fluorescent visible implant elastomer (VIE) tagging system in juvenile gilthead seabream, Sparus auratus L., between 7 and 18 g was examined. Four different colours (red, green, orange and yellow), three body positions (dorsal, lateral and caudal) and two orientations (horizontal and vertical) were tested. The mean tag application time for each fish was 15.7±0.32 s. There was no mortality associated with the method of tagging. The most visible tag colour was red. Injection orientation had a significant effect on length, width, fragmentation and fluorescent intensity of the tag. Horizontal tagging is recommended because of high fluorescent intensity, low fragmentation and for double tagging. There were no differences in growth between untagged controls and the VIE‐tagged fish. 相似文献
155.
Abstract Growth and total mortality of Australian bass, Macquaria novemaculeata Steindachner, were studied in four freshwater impoundments using data from a cooperative angler tagging programme. Growth was modelled using the computer program GROTAG. Annual growth increments varied among impoundments, ranging from 5.0 to 7.8 cm yr−1 among fish 20-cm total length (TL) and 1.7 to 4.9 cm yr−1 among fish 30 cm TL, but exceeded previous estimates for the species. Annual total mortality rates, based on the decrease in recapture rate with time, ranged from 0.03 to 0.07 and were lower than for other comparable species, presumably because of the prevalence of catch and release fishing in the impoundments studied. 相似文献
156.
157.
158.
Surinder Chopra Amanda Gevens Catherine Svabek Karl V. Wood Thomas Peterson Ralph L. Nicholson 《Physiological and Molecular Plant Pathology》2002,60(6):321
The 3-deoxyanthocyanidin phytoalexins produced in sorghum leaves in response to Colletotrichum sublineolum have chemical structure similarities to the 3-deoxy flavonoids that are precursors of phlobaphene pigments. Phlobaphenes are commonly observed in the pericarp of mature sorghum grains, while synthesis of 3-deoxyanthocyanidin phytoalexins is a site-specific response to infection with C. sublineolum. We have taken a genetic approach to investigate the possible overlap between the two sub-branches of flavonoid biosynthesis in sorghum that lead to phlobaphenes and 3-deoxyanthocyanidin phytoalexins. A sorghum line with a functional y1 gene synthesizes 3-deoxyanthocyanidins as well as phlobaphenes. However, a progenitor line with the mutable Y1-candystripe (Y1-cs) allele shows variable levels of biosynthesis of these compounds. The Y1-cs allele carries a copy of the Candystripe1 (Cs1) transposable element in the y1 gene. We demonstrate here that the variability in the expression of 3-deoxyanthocyanidins produced in individual mesocotyls of hyper-mutable Y1-cs plants is a function of the activity of the y1 gene. TheCs1 insertion in the Y1-cs allele blocks y1 function, while excision of Cs1 out of they1 locus restores the gene to a functional state. The combined molecular and biochemical characterization of sibling plants confirms that the allelic state of the y1 gene is completely correlated with the production of phytoalexins in response to fungal infection. These results provide support for the idea that the y1 gene regulates the biosynthesis of both 3-deoxyanthocyanidin phytoalexins and phlobaphene pigments in sorghum. 相似文献
159.
Rieko Hatta Akihisa Shinjo Sheila Ruswandi Keiji Kitani Mikihiro Yamamoto Kazuya Akimitsu Takashi Tsuge 《Journal of General Plant Pathology》2006,72(4):210-219
The strawberry pathotype of Alternaria alternata produces host-specific AF-toxin and causes Alternaria black spot of strawberry. Previously, we isolated cosmid clones pcAFT-1
and pcAFT-2 from strain NAF8 of the strawberry pathotype that contain AF-toxin biosynthetic genes, named AFT genes. In a molecular characterization here of pcAFT-1 and pcAFT-2, 11 AFT genes and five transposon-like sequences, named TLS-S1 to TLS-S5, were detected. The nucleotide sequences of TLS-S1 and TLS-S4
share high homology, and their putative products have similarity to transposases of the hAT family transposons. Thus, TLS-S1 and TLS-S4 were renamed TLS-S1-1 and TLS-S1-2, respectively. Amino acid sequences deduced
from TLS-S2, TLS-S3, and TLS-S5 have similarity to transposases of the Fot1/Pogo family transposons, but they are significantly different. All five sequences have incomplete open reading frames (ORFs) for
transposases owing to deletions, termination codons, and/or frameshifts, indicating that they are inactivated elements. Analysis
of genomic distribution of these sequences revealed that they are specifically distributed on a 1.05-Mb chromosome of NAF8,
which has been identified as a conditionally dispensable (CD) chromosome encoding AFT genes. The presence of three, four, and three copies of TLS-S1, TLS-S2, and TLS-S3, respectively, and a single copy of TLS-S5
on the CD chromosome were estimated by DNA gel blot analysis. The remaining copy of TLS-S1 and the three copies of TLS-S2
were isolated and identified to also encode incomplete ORFs. Thus, it appears that all copies of the transposon-like sequences
identified are inactivated elements (fossils) unique to the CD chromosome in the genome of the strawberry pathotype.
The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB236733
(TLS-S1-1), AB236734 (TLS-S1-2), AB236735 (TLS-S1-3), AB236736 (TLS-S2-1), AB236737 (TLS-S2-2), AB236738 (TLS-S3), and AB236739
(TLS-S5) 相似文献
160.
为在大肠杆菌全基因组范围内筛选和鉴定与喹诺酮类抗生素耐药性相关的基因,通过对临床分离的13株大肠杆菌进行药敏试验,选择耐药谱较广泛的菌株作为研究对象,利用Mariner转座子对其基因组进行随机突变,获得转座子突变株文库,并以亲本菌株为对照筛选文库中对喹诺酮类抗生素敏感的突变体,通过套式PCR、核苷酸测序及序列比对确定突变株中转座子的插入位点及其破坏的基因。结果表明:从突变株文库中筛选得到22株分别对诺氟沙星、环丙沙星、左氧氟沙星、萘啶酸敏感的突变株,其中7株对4种抗生素都敏感;插入位点分析发现,抗生素敏感突变株中被转座子破坏的基因包括ecs4206(磷酸核酮糖激酶)、ecs3959(β-D-半乳糖苷酶β亚基)、ecs3946(假定蛋白)和ecs1857(DNA结合转录调控因子)。筛选出的基因可被开发为控制或扭转大肠杆菌耐药性的作用靶点。 相似文献