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Cloning of rainbow trout (Oncorhynchus mykiss) {alpha}-actin, myosin regulatory light chain genes and the 5'-flanking region of {alpha}-tropomyosin. Functional assessment of promoters

Aleksei Krasnov1,*, Heli Teerijoki1, Yuri Gorodilov2 and Hannu Mölsä1

1 Institute of Applied Biotechnology, University of Kuopio, P.O.B. 1627, FIN-Kuopio 70211, Finland
2 Biological Institute, University of Sanct Petersburg, Oranienbaum Chaussee 2, Stary Peterhof, Sanct Petersburg 198504, Russia



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  		Fig. 1. Map of construct including {alpha}-OnmyAct promoter, the coding part of OnmyGLUT1 and {alpha}-OnmyAct terminator. Location of PCR primers is indicated by arrows.

 


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  		Fig. 2. Rainbow trout {alpha}-actin ({alpha}-OnmyAct). (A) Exon structure of vertebrate {alpha}-actin genes: rainbow trout (this study), pufferfish skeletal 2 (GenBank U38958) and cardiac 1-3 (GenBank U38959, U38960, U38961), and chick cardiac (GenBank X02212). (B) Putative binding sites of transcription regulatory factors in the promoter region and first exon.

 


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  		Fig. 3. Rainbow trout regulatory myosin light chain (OnmyMLC2). (A) Two promoters include a conserved 800-bp sequence upstream from the transcription start and regions similar to different fish transposon pseudogenes; arrows show orientation. Putative TATA and E-boxes are indicated. (B) Exon structure of rainbow trout (this study) and rat muscle (GenBank X00975) genes.

 


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  		Fig. 4. Expression of {alpha}-OnmyTMLacZ in rainbow trout embryo at the stage of 39 somite pairs. (A) LacZ-positive cells were detected in somites (s) and the neural plate (np). (B) Reporter expression was seen in the neural crest (nc) on the border between the neural plate (np) and presomite mesoderm (psm). s, somite. (C) LacZ was expressed in mesencephalon (me) and neuromeres of the hindbrain (nm). mt, myotome; ysl, yolk syncytial layer. (D) Heart (h). (E) LacZ-positive cells were detected in the rest of the yolk syncytial layer (ysl).

 

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© The Company of Biologists Ltd 2003