Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81

Warning: fopen(upload/ip_log/ip_log_2024-03.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83

Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84
Genetic comparison between Spirometra erinacei and S. mansonoides using PCR-RFLP analysis
| Home | E-Submission | Sitemap | Contact us |  
top_img
Korean J Parasito Search

CLOSE

Korean J Parasito > Volume 35(4):1997 > Article

Original Article
Korean J Parasitol. 1997 Dec;35(4):277-282. English.
Published online Dec 20, 1997.  http://dx.doi.org/10.3347/kjp.1997.35.4.277
Copyright © 1997 by The Korean Society for Parasitology
Genetic comparison between Spirometra erinacei and S. mansonoides using PCR-RFLP analysis
S U Lee,1S Huh,*1 and C K Phares2
1Department of Parasitology, College of Medicine, Hallym University, Chunchon 200-702, Korea.
Received October 13, 1997; Accepted November 20, 1997.

Abstract

The only observed morphological difference between Spirometra erinacei and S. mansonoides is the uterine shape of the mature proglottid. Two species of worms are thought to be evolutionarily closely related. Biomolecular comparison of the two worms by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was conducted to observe the genetic distance. The 28s rDNA, mitochondrial cytochrome c oxidase subunit I (mCOI), and ribosomal internal transcribed spacer 1 (ITS1) fragments were obtained from the worms by PCR. The PCR products were cleaved by 5 four-base pair restriction enzyme combinations (Msp I, Hae III, Alu I, Cfo I, Rsa I), electrophoresed and analyzed with PAUP 3.1.1. The fragment patterns of 28S rDNA and ITS1 demonstrated that two worms were in identical systematic tree with bootstrap number 94 and 100, respectively. As for mCOI, bootstrap number was 74 in a different tree. Above results are indicative of recent common ancestry between S. erinacei and S. mansonoides.

Figures


Fig. 1
Two percent agarose gel electrophoresis of PCR-RFLP for 28S rDNA D1 using the restriction endonuclease. Lane M, molecular weight markers; lane 1, S. erinacei; lane 2, S. mansonoides; lane 3, T. asiatica; lane 4, T. taeniaeformis; lane 5, H. diminuta; lane 6, D. latum.


Fig. 2
Two percent agarose gel electrophoresis of PCR-RFLP for mCOI using the restriction endonuclease. Lane M, molecular weight markers; lane 1, S. erinacei; lane 2, S. mansonoides; lane 3, T. asiatica; lane 4, T. taeniaeformis; lane 5, H. diminuta; lane 6, D. latum.


Fig. 3
Two percent agarose gel electrophoresis of PCR-RFLP for rDNA ITS1 using the restriction endonuclease. Lane M, molecular weight markers; lane 1, S. erinacei; lane 2, S. mansonoides; lane 3, T. asiatica; lane 4, T. taeniaeformis; lane 5, H. diminuta; lane 6, D. latum.


Fig. 4
Molecular phylogenetic analyses of PCR-RFLP obtained using 28S rDNA, mCOI and the rDNA ITS1 gene. Branch lengths and ranges of bootstrap values were obtained using statistical package (Power Macintosh 6100/66, MacClade V 3.0., Clustal V and PAUP Ver.3.1.1). Bootstrap values are given as a percent of the above corresponding branch. A, 28S rDNA; mCOI; C, rDNA ITS1 gene.

References
1. Bowditch BM, Albright DG, Williams JG, Braun MJ. Use of randomly amplified polymorphic DNA markers in comparative genome studies. Methods Enzymol 1993;224:294–309.
  
2. Bowles J, McManus DP. Rapid discrimination of Echinococcus species and strains using a polymerase chain reaction-based RFLP method. Mol Biochem Parasitol 1993;57(2):231–239.
  
3. Bowles J, McManus DP. Genetic characterization of the Asian Taenia, a newly described taeniid cestode of humans. Am J Trop Med Hyg 1994;50(1):33–44.
 
4. Fukumoto S, Tsuboi T, Hirai K, Phares CK. Comparison of isozyme patterns between Spirometra erinacei and Spirometra mansonoides by isoelectric focusing. J Parasitol 1992;78(4):735–738.
  
5. Garey JR, Wolstenholme DR. Platyhelminth mitochondrial DNA: evidence for early evolutionary origin of a tRNA(serAGN) that contains a dihydrouridine arm replacement loop, and of serine-specifying AGA and AGG codons. J Mol Evol 1989;28(5):374–387.
  
6. Higgns DG, et al. Computer of Applied Bioscience 1994;8:189–191.
8. Iwata S. Prog Med Parasit in Japan 1972;4:536–590.
9. Lee SH, Chai JY, Seo BS, Cho SY. Two cases of human infection by adult of Spirometra erinacei. Korean J Parasitol 1984;22(1):66–71.
 
11. Mueller JF. The biology of Spirometra. J Parasitol 1974;60(1):3–14.
 
12. Orita M, Suzuki Y, Sekiya T, Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 1989;5(4):874–879.
  
13. Qu LH, Nicoloso M, Bachellerie JP. Phylogenetic calibration of the 5' terminal domain of large rRNA achieved by determining twenty eucaryotic sequences. J Mol Evol 1988;28(1-2):113–124.
  
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2024 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI