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林麝染色体制备方法及核型与G- 带带型研究

邹方东  岳碧松  张义正  陈三   

  1. 四川大学生命科学学院
  • 出版日期:2005-07-14 发布日期:2008-07-07

Studies on the Karyotype and Chromosomal G2Banding of

ZOU Fangdong YUE Bisong ZHANG Yizheng CHEN San   

  • Online:2005-07-14 Published:2008-07-07

摘要: 以林麝外周血淋巴细胞为实验材料, 在培养基中加入植物血球凝集素PHA 和伴刀豆球蛋白ConA , 由此建立了适合其增殖的培养体系。培养75 h 后, 用空气干燥法制备染色体, 确定林麝核型是2N = 58 , 且全都是端着丝粒染色体。实验结果还表明, 通过获得较晚的中期分裂相, 可以确定染色体是端着丝粒不是亚端着丝粒类型。同时首次应用染色体G- 带技术, 研究了林麝染色体的G- 带带型。

关键词: 林麝, 核型, 端着丝粒染色体, G- 带

Abstract: A unique culture system has been developed to induce more peripheral blood lymphocytes of musk deer (Moschus berezovskii) into cell cycle by adding an equal quantity of phytohemagglutinin (PHA) and concanavalin A (ConA) , which stimulate G0 phase lymphocytes to proliferate. Peripheral blood samples from 5 musk deers (3 ♂、2 ♀) of Institute of Scihuan Musk Deer Breeding was obtained from ear vein with heparin as an anticoagulant , and inoculated 0.3 - 0.4 ml of blood into 54 ml of RPMI 1640 medium containing 1 ml of bovine serum , 40μg of PHA and ConA , 0.05 ml of heparin (10 mg/ ml) and an appropriate antibiotics. The pH value of medium was adjusted to 7.0 - 7.21 The blood was cultured for 75 hrs and added colchicine to a final concentration of 0.01μg/ ml before the end of culturing. Preparation of chromosome was carried out with air-dry method and displayed G-bands with 0.025 % trypsin. It was found that the karyotype of M. berezovskii was 2N = 58 , and all of the chromosomes are telocentric ones. The results indicated that using late metaphases , in which all the chromosomes are linked only by centromeres , can determine whether the chromosome is telocentric or not. This study was the first time to use G-banding technique to analyze the chromosomes of M. moschiferus.

Key words: Musk deer (Moschus moschiferus), Karyotype, Telocentric chromosomes, G-banding