Fluorescent in situ hybridization on mitotic chromosomes of mosquitoes.
Fluorescent in situ hybridization on mitotic chromosomes of mosquitoes.
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DOI：
10.3791/4215
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发表时间：
2012-09-17
期刊：
影响因子：
--
通讯作者：
Sharakhova MV
中科院分区：
文献类型：
--
作者：
Timoshevskiy VA;Sharma A;Sharakhov IV;Sharakhova MV
荧光原位杂交（FISH）是许多实验室常规使用的技术，用于确定DNA和RNA探针的染色体位置。该方法的一个重要应用是开发高质量的物理图谱，有助于改善各种生物体的基因组组装。多丝和有丝分裂染色体的自然带带模式为基因组超contigs的精确排序和方向提供了指导。在按蚊、伊蚊和库蚊这三种蚊子属中，一种完善的基于染色体的定位技术仅用于按蚊，其成员具有可读的多烯染色体1。由于基因组图谱的努力，88%的印第安人。冈比亚基因组被精确地放置在染色体2,3的位置上。另外两种蚊属，伊蚊和库蚊，由于其基因组中转座因子的显著过度表达，染色体多聚化程度较低4,5,6。只有31%和9%的基因组超序列被无顺序或定向地分配到伊蚊的染色体上。埃及伊蚊7和Cx。分别为致倦库蚊8只。这两个物种的有丝分裂染色体制备以前仅限于脑神经节和细胞系。然而，从蚊子的大脑神经节制备的染色体载玻片通常含有少量的中期板。此外，尽管FISH技术已经发展为有丝分裂染色体的Ae细胞系。埃及伊蚊，细胞系染色体中多染色体重排的积累使得它们对基因组绘制毫无用处。本文描述了一种简单、可靠的技术，从4龄幼虫的成像盘（IDs）中获得高质量的有丝分裂染色体，可用于所有三属蚊子。一种标准的FISH方案12被优化，用于使用基因组DNA的BAC克隆作为伊蚊有丝分裂染色体的探针。埃及伊蚊和Cx。利用核糖体DNA （rDNA）的一个基因间间隔区（IGS）作为一种探针。冈比亚按蚊的染色体。除物理制图外，所开发的技术还可用于蚊子和其他昆虫类群的群体细胞遗传学和染色体分类/系统分类。
Fluorescent in situ hybridization (FISH) is a technique routinely used by many laboratories to determine the chromosomal position of DNA and RNA probes. One important application of this method is the development of high-quality physical maps useful for improving the genome assemblies for various organisms. The natural banding pattern of polytene and mitotic chromosomes provides guidance for the precise ordering and orientation of the genomic supercontigs. Among the three mosquito genera, namely Anopheles, Aedes, and Culex, a well-established chromosome-based mapping technique has been developed only for Anopheles, whose members possess readable polytene chromosomes 1. As a result of genome mapping efforts, 88% of the An. gambiae genome has been placed to precise chromosome positions 2,3 . Two other mosquito genera, Aedes and Culex, have poorly polytenized chromosomes because of significant overrepresentation of transposable elements in their genomes 4, 5, 6. Only 31 and 9% of the genomic supercontings have been assigned without order or orientation to chromosomes of Ae. aegypti 7 and Cx. quinquefasciatus 8, respectively. Mitotic chromosome preparation for these two species had previously been limited to brain ganglia and cell lines. However, chromosome slides prepared from the brain ganglia of mosquitoes usually contain low numbers of metaphase plates 9. Also, although a FISH technique has been developed for mitotic chromosomes from a cell line of Ae. aegypti 10, the accumulation of multiple chromosomal rearrangements in cell line chromosomes 11 makes them useless for genome mapping. Here we describe a simple, robust technique for obtaining high-quality mitotic chromosome preparations from imaginal discs (IDs) of 4th instar larvae which can be used for all three genera of mosquitoes. A standard FISH protocol 12 is optimized for using BAC clones of genomic DNA as a probe on mitotic chromosomes of Ae. aegypti and Cx. quinquefasciatus, and for utilizing an intergenic spacer (IGS) region of ribosomal DNA (rDNA) as a probe on An. gambiae chromosomes. In addition to physical mapping, the developed technique can be applied to population cytogenetics and chromosome taxonomy/systematics of mosquitoes and other insect groups.
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DOI：
10.1007/978-1-60761-789-1_1
发表时间：
2010-01-01
期刊：
FLUORESCENCE IN SITU HYBRIDIZATION (FISH): PROTOCOLS AND APPLICATIONS
影响因子：
--
作者：
Garimberti, Elisa;Tosi, Sabrina
通讯作者：
Tosi, Sabrina
DOI：
10.1139/g75-031
发表时间：
1975-01-01
期刊：
CANADIAN JOURNAL OF GENETICS AND CYTOLOGY
影响因子：
--
作者：
STEINIGER, GE;MUKHERJEE, AB
通讯作者：
MUKHERJEE, AB
影响因子：
3.8
作者：
Sharakhova MV;Timoshevskiy VA;Yang F;Demin SIu;Severson DW;Sharakhov IV
通讯作者：
Sharakhov IV
影响因子：
2.1
作者：
Dasgupta, Ranjit;Free, Heather M.;Christensen, Bruce M.
通讯作者：
Christensen, Bruce M.
DOI：
10.1126/science.1191864
发表时间：
2010-10-01
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Arensburger P;Megy K;Waterhouse RM;Abrudan J;Amedeo P;Antelo B;Bartholomay L;Bidwell S;Caler E;Camara F;Campbell CL;Campbell KS;Casola C;Castro MT;Chandramouliswaran I;Chapman SB;Christley S;Costas J;Eisenstadt E;Feschotte C;Fraser-Liggett C;Guigo R;Haas B;Hammond M;Hansson BS;Hemingway J;Hill SR;Howarth C;Ignell R;Kennedy RC;Kodira CD;Lobo NF;Mao C;Mayhew G;Michel K;Mori A;Liu N;Naveira H;Nene V;Nguyen N;Pearson MD;Pritham EJ;Puiu D;Qi Y;Ranson H;Ribeiro JM;Roberston HM;Severson DW;Shumway M;Stanke M;Strausberg RL;Sun C;Sutton G;Tu ZJ;Tubio JM;Unger MF;Vanlandingham DL;Vilella AJ;White O;White JR;Wondji CS;Wortman J;Zdobnov EM;Birren B;Christensen BM;Collins FH;Cornel A;Dimopoulos G;Hannick LI;Higgs S;Lanzaro GC;Lawson D;Lee NH;Muskavitch MA;Raikhel AS;Atkinson PW
通讯作者：
Atkinson PW