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Korean J Parasitol > Volume 30(3):1992 > Article

Original Article
Korean J Parasitol. 1992 Sep;30(3):183-189. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1992.30.3.183
Copyright © 1992 by The Korean Society for Parasitology
Karyotypes of Pneumocystis carinii from Korean rats
Sung-Tae Hong,1Beyong Il Kim,2Weon-Gyu Kho,1,*Jae-Ran Yu,1,*Jina Kook,1Jong-Yil Chai,1Chong-Ku Yun,2 and Soon-Hyung Lee1
1Department of Parasitology, Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea.
1Department of Pediatrics, Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea.

**Weon-Gyu Kho, Department of Parasitology, Inje University, Pusan 614-735; Jea-Ran Yu, Department of Parasitology, College of Medicine, Konkuk University, Chungju 380-701.

Abstract

Molecular karyotyping was applied to Pneumocystis carinii(Pc) from two strains of experimental rats, Sprague Dawley(SD) and Fisher(F), in Korea. Field inversion gel electrophoresis and contour clamped homogeneous electric field electrophoresis resolved 15 chromosomal bands from the Pc. The size of the bands was estimated 270kb to 684kb from SD rats, and 273kb to 713 kb from F rats. The bands of 283 kb from SD rats and of 273 kb from F rats stained more brightly suggesting duplicated bands. Total number of chromosomes was at least 16, and total genomic size was estimated 7 × 106 bp. All of the bands from F rats hybridized to the probe of repeated DNA sequences of Pc and the band of 448 kb size was proved to contain rDNA sequences, but Pc. chromosome bands from SD rats showed no reactions to the probes. The 2 different karyotypes of P. carinii from 2 strains of rats were maintained consistently for 2 years.

Figures


Fig. 1
A: CHEF gel (C15) run in 1% agarose under 95 V for 120 hrs, initial A time 25 sec, final A time 150 sec, A/B ratio 1, 1/2X TBE buffer at 15℃. Lanes loaded with size marker and Pneumocystis carinii (Pc) plugs. 1. Saccharomyces cerevisiae AB 972, a size marker; 2. USD6-2, Pc from Sprague Dawley(SD) rats in the USA; 3. UL8-5, Pc from Lewis rats in the USA; 4. SD5-1, Pc from SD rats (1/24/91); 5. SD8-2, Pc from SD rats (4/12/91); 6. F14, Pc from Fisher(F) rats (12/31/90); 7. F15, Pc from F rats (7/12/91). B: Autoradiograph after hybridization of P32 labeled Rp3-1 probe to the Southern blot of the gel Fig. 1A (C15), exposed to the film after high stringency washing. The lanes 2(USD6-2), 3(UL8-5), 6(F14) and 7(F15) showed strong signals in all bands suggesting that they share common repeat sequences of Pc.


Fig. 2
A: CHEF gel (C13) of 0.8% agarose run under conditions of 105 V, 72 hrs, initial A time 25 sec, final A time 110 sec, and 1/2X TBE buffer at 15℃. Loaded plugs are 1. Size marker Saccharomyces cerevisiae AB 972; 2. USD5-2, Pc from SD rats in the USA; 3. SD3, Pc from SD rats (11/13/90); 4. SD5-2, Pc from SD rats (1/24/19); 5. SD8-1, Pc from SD rats (4/12/91); 6. SD8-2, Pc from SD rats (4/12/91); 7. SD12, Pc from SD rats (8/20/91); 8. F1-1, Pc from F rats (3/26/90); 9. F1-3, Pc from F rats (4/2/90); 10. F14, Pc from F rats (12/31/90); 11. F15, Pc from F rats (7/12/91). B: Autoradiograph of hybridization of P32 labeled rat DNA probe to the blot of Fig. 2A (C13) after washing in high stringency solution. Intense signals are observed on the plugs in the trough, megabase-sized bands, and smear backgrounds. The resolved bands between 270 kb and 700 kb show no reactions. Also the reactions differ by individual samples.


Fig. 3
A: CHEF gel (C14) of 1% agarose run under the parameters 90 V, 96 hrs, initial A time 25 sec, final A time 120 sec, A/B ratio 1, 1/2X TBE buffer at 15℃. The samples loaded are 1. Saccharomyces cerevisiae AB 972; 2. USD6-2, Pc from SD rats in the USA; 3. SD8-2, Pc from SD rats (4/12/91); 4. F15, Pc from F rats (7/12/91). B: Autoradiograph of the Fig. 3A (C14) gel blot, hybridized to the prove of 19c-1 rRNA genomic DNA. The 448 kb band of F15 reacted strongly. The bands of the yeast and one band of USD6-2 also showed signals.


Fig. 4
The FIGE gel, F30, run under conditions of 1% agarose, 50 sec forward and 25 sec backward, 105 V, 96 hrs, and 1/2X TBE buffer 14℃. Loaded samples are 1. Size marker Saccharomyces cerevisiae AB 972; 2. SD8-1; 3. F14; 4. SD8-1; 5. F15; 6. SD8-2; 7. F14; 8. SD8-2; 9. F15.


Fig. 5
Estimated size of chromosomal molecular bands resolved from SD8-1 and F15. The 283 kb band of SD8-1 and the 273 kb band of F15 are suggestive of doublet of 2 or more bands of same size.

Tables


Table 1
List of Pc sample from albino rats

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