3988 Nucleic Acids Research, Vol. 18, No. 13

Sequence

of

a

tRNAGIY from Streptomyces coelicolor

J.Stefan Rokem*, Astrid Sch6n+ and Dieter So6I Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 0651 1, USA EMBL accession

Submitted June 5, 1990

METHODS

no.

X51702

A C

was isolated from S. coelicolor A3 (2) J650. The cells were grown for 48 h at 30°C and 150 rpm in AM medium (1), broken by N2 cavitation (2) and the lysate was passed directly into phenol, extracted and precipitated. tRNAs were deacylated, purified on DEAE cellulose and separated by RPC-5 chromatography (3). End labeled tRNAs were purified to homogeneity on a 20% polyacrylamide/8M urea gel, and

tRNA

C U

pG0-C C -G G -C -C

G-c U

sequenced by standard methods (4, 5).

U

AA U

RESULTS We have isolated and sequenced a tRNA from Streptomyces coelicolor with a glycine specific anticodon, CCC. The tRNA shows a high level of homology to tRNAGIY(CCC) and tRNAGlY(UCC) from E. coli (81 and 74%), a tRNAGlY(UCC) gene from Pseudomonas aeruginosa (86%), and a tRNAGIY(UCC) from B. subtilis (86%)

REFERENCES

-A

G-C G

CUUG

A

UGCUC .*,,, GCGAG

I

uu

A

G

C T4

GGDAGA D AGAAC

G

G

U -AG C -G A -U G-C C -G A U U A c

A

c

Sequence of tRNA Gl from S. coelicolor A3(2)

1. Feitelson,J.S., Malpartida,F. and Hopwood,D.A. (1985) J. Gen. Microbiol. 131, 2431-2441. 2. Osmani,S.A., Marston,F.A.O., Selmes,I.P., Chapman,A.G. and Scrutton,M.C. (1981) Eur. J. Biochem. 118, 271-278. 3. Pearson,R.L., Weiss,J.F. and Kelmers,A.D. (1971) Biochlm. Biophys. Acta 228, 770-774. 4. Krupp,G. and Gross,H.J. (1983) In 7he Modified Nucleotides in Transfer RN/A I. Liss, New York, 11-58. 5. Waldmann,R., Gross,H.J. and Krupp,G. (1987) Nucl. Acids Res. 15, 7209. 6. Sprinzl,M., Hartmann,T., Weber,J., Blank,J. and Zeidler,R. (1989) Nucl. Acids Res. 17, rl -r172.

Present addresses: *Department of Applied Microbiology, Hebrew University, Jerusalem, Israel and +Laboratorium fiir Biochemie,

Bayreuth, FRG

Universitiit Bayreuth,

8580