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《Acta Zoologica Sinica》 2002-01
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MICROISOLATION AND CHARACTERIZATION OF SINGLE CHROMOSOME IN RICE FIELD EELS (MONOPTERUS ALBUS)

JI Fu-Yun YU Qi-Xing **- GUO Yi-Qing HAI Yan\ ZHOU Rong-Jia\ LIU Li (College of Life Science, Wuhan University, Wuhan 430072, China)  
The methods of microisolation and characteristic assay of single chromosome in rice field eels (Monopterus albus) were established. The chromosome 3 of meiosis Ⅰ diakinesis was microisolated manually with a micro-glass needle under the Olympus reverted microscope and its DNA was amplified by DOP-PCR. Several hybridizations were carried out to detect its specialities. 18 S rRNA gene of soybean labeled by Biotin-11-dUTP were hybridized with the metaphase chromosome and the DOP-PCR products dotted on the nylon membrane to map rice field eel rRNA genes.The DOP-PCR products with the positive Dot hybridization results labeled by Biotin-11-dUTP and α-32P-dCTP respectively were hybridized with the genomic DNA digested by different enzymes and the metaphase chromosome to further confirm its specialities. The hybridization results show that: (1) Chromosome specimen of meiosis Ⅰ diakinesis is the ideal material for single chromosome microisolation; (2) The sizes of DOP-PCR products range from 200 bp to 1 000 bp, averagely 600 bp;(3) The single chromosome obtained in the study is rice field eel chromosome 3;(4) Compared with micro-manipulator and micro-laser beam, the methods developed in the study can be more widely used in ordinary laboratory for requiring no expensive instrument.In the paper, we also discuss the future application of microisolation techniques in the study of chromosome evolution, gene mapping and conserved synteny in fish.
【Fund】: 国家自然科学基金资助项目 (No .39970 40 6 );; 高等学校博士学科点专项科研基金资助项目 (No.980 486 2 8)~~
【CateGory Index】: Q959.46
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