Identification, cloning, and characterization of microsatellite DNA in Euglena gracilis.
Microsatellite DNA has been developed into one of the most popular genetic markers. We have identified and cloned microsatellite loci in the genome of a free-living protozoan Euglena gracilis FACHB-848, using the random amplified microsatellites method (RAMS). The digoxigenin-labelled oligonucleotides (CT)10 and (GT)10 served as probes to detect complementary sequences in the randomly amplified polymorphic DNA (RAPD) fingerprints produced by means of Southern blotting. Subsequently, positive RAPD fragments were cloned. From a total of 31 RAPD primer profiles, eight microsatellite loci of E. gracilis were detected and characterized. Further, six sites (i.e. EGMS1, EGMS3, EGMS4, EGMS5, EGMS6, and EGMS7) showed polymorphisms. We found a GT or CT microsatellite every 10.5 kb in the genome of E. gracilis, and similar to animal genomes, the (GT)(n) motif was much more abundant than the (CT)(n) motif. These polymorphic microsatellite DNA will serve as advantageous molecular markers for studying the genetic diversity and molecular ecology of Euglena.
Zhang
Wen-Jing
WJ
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobioilogy, Chinese Academy of Sciences, Wuhan 430072, China.
Yang
Jun
J
Yu
Yu-He
YH
Shen
Yun-Fen
YF
D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D005056:Euglena gracilis / Q000235:genetics*; D017403:In Situ Hybridization; D018895:Microsatellite Repeats / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA
D016054:DNA, Protozoan