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Plant Physiol, January 2001, Vol. 125, pp. 430-436
Sequences Downstream of the Translation Initiation Codon Are
Important Determinants of Translation Efficiency in
Chloroplasts1
Hiroshi
Kuroda and
Pal
Maliga*
Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen Road, Piscataway, New Jersey 08854-8020
The objective of this study was to determine if mRNA sequences
downstream of the translation initiation codon are important for
translation of plastid mRNAs. We have employed a transgenic approach,
measuring accumulation of the neomycin phosphotransferase (NPTII)
reporter enzyme translationally fused with 14 N-terminal amino acids
encoded in the rbcL or atpB plastid
genes. NPTII accumulation from wild-type and mutant rbcL
and atpB segments was compared. We report that silent
mutations in the rbcL segment reduced NPTII accumulation
35-fold. In contrast, mutations in the atpB mRNA reduced
NPTII accumulation only moderately from approximately 7% (w/w) to
approximately 4% (w/w) of the total soluble cellular protein,
indicating that the importance of sequences downstream of the
translation initiation codon are dependent on the individual mRNA.
Information provided here will facilitate transgene design for
high-level expression of recombinant proteins in chloroplasts by
translational fusion with the N-terminal segment of highly expressed
plastid genes or by introduction of silent mutations in the N-terminal
part of the coding region.
1
This research was supported by the National
Science Foundation (grant no. MCB 96-30763).
*
Corresponding author; e-mail maliga{at}waksman.rutgers.edu; fax
732-445-5735.
© 2001 American Society of Plant Physiologists
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