林麝染色体制备方法及核型与G-带带型研究

Studies on the Karyotype and Chromosomal G-Banding of Musk Deer(Moschus berezovskii)with a New Method

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75h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。

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 G0 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, 40 μg of PHA and ConA, 0.05 ml of heparin (10 mg/ml) and an appropriate antibiotics. The pH value of medium was adjusted to 7.0-7.2. The blood was cultured for 75 hrs and added colchicine to a final concentration of 0.01 μg/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.