Dataset Information

Nuclear-encoded chloroplast ribosomal protein L12 of Nicotiana tabacum: characterization of mature protein and isolation and sequence analysis of cDNA clones encoding its cytoplasmic precursor.

ABSTRACT: Poly(A)+ mRNA isolated from Nicotiana tabacum (cv. Petite Havana) leaves was used to prepare a cDNA library in the expression vector lambda gt11. Recombinant phage containing cDNAs coding for chloroplast ribosomal protein L12 were identified and sequenced. Mature tobacco L12 protein has 44% amino acid identity with ribosomal protein L7/L12 of Escherichia coli. The longest L12 cDNA (733 nucleotides) codes for a 13,823 molecular weight polypeptide with a transit peptide of 53 amino acids and a mature protein of 133 amino acids. The transit peptide and mature protein share 43% and 79% amino acid identity, respectively, with corresponding regions of spinach chloroplast ribosomal protein L12. The predicted amino terminus of the mature protein was confirmed by partial sequence analysis of HPLC-purified tobacco chloroplast ribosomal protein L12. A single L12 mRNA of about 0.8 kb was detected by hybridization of L12 cDNA to poly(A)+ and total leaf RNA. Hybridization patterns of restriction fragments of tobacco genomic DNA probed with the L12 cDNA suggested the existence of more than one gene for ribosomal protein L12. Characterization of a second cDNA with an identical L12 coding sequence but a different 3'-noncoding sequence provided evidence that at least two L12 genes are expressed in tobacco.

SUBMITTER: Elhag GA

PROVIDER: S-EPMC312006 | biostudies-other | 1992 Feb

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

Nuclear-encoded chloroplast ribosomal protein L12 of Nicotiana tabacum: characterization of mature protein and isolation and sequence analysis of cDNA clones encoding its cytoplasmic precursor.

Elhag G A GA Thomas F J FJ McCreery T P TP Bourque D P DP

Nucleic acids research 19920201 4

Poly(A)+ mRNA isolated from Nicotiana tabacum (cv. Petite Havana) leaves was used to prepare a cDNA library in the expression vector lambda gt11. Recombinant phage containing cDNAs coding for chloroplast ribosomal protein L12 were identified and sequenced. Mature tobacco L12 protein has 44% amino acid identity with ribosomal protein L7/L12 of Escherichia coli. The longest L12 cDNA (733 nucleotides) codes for a 13,823 molecular weight polypeptide with a transit peptide of 53 amino acids and a mat ...[more]

PMID: 1542565

Similar Datasets

Project description:BackgroundChloroplast is indispensable for plant response to environmental stresses, growth and development, whose function is regulated by different plant hormones. The chloroplast proteome is encoded by chloroplast genome and nuclear genome, which play essential roles in plant photosynthesis, metabolism and other biological processes. Ethylene response factors (ERFs) are key transcription factors in activating the ethylene signaling pathway and plant response to abiotic stress. But we know little about how ethylene regulates plastid function under drought stress condition. In this study we utilized tobacco overexpressing tomato ethylene responsive factor 1 (TERF1), an ERF transcription factor isolated from tomato, to investigate its effects on the plastid proteome under drought stress condition by method of iTRAQ technology.ResultsResults show that TERF1 represses the genes encoding the photosynthetic apparatus at both transcriptional and translational level, but the genes involved in carbon fixation are significantly induced by TERF1. TERF1 regulates multiple retrograde signaling pathways, providing a new mechanism for regulating nuclear gene expression. TERF1 also regulates plant utilization of phosphorus (Pi) and nitrogen (N). We find that several metabolic and signaling pathways related with Pi are significantly repressed and gene expression analysis shows that TERF1 significantly represses the Pi transport from root to shoot. However, the N metabolism is upregulated by TERF1 as shown by the activation of different amino acids biosynthesis pathways due to the induction of glutamine synthetase and stabilization of nitrate reductase although the root-to-shoot N transport is also reduced. TERF1 also regulates other core metabolic pathways and secondary metabolic pathways that are important for plant growth, development and response to environmental stresses. Gene set linkage analysis was applied for the upregulated proteins by TERF1, showing some new potential for regulating plant response to drought stress by TERF1.ConclusionsOur research reveals effects of ethylene signaling on plastid proteome related with two key biological processes, including photosynthesis and nutrition utilization. We also provide a new mechanism to regulate nuclear gene expression by ERF1 transcription factor through retrograde signals in chloroplast. These results can enrich our knowledge about ERF1 transcription factor and function of ethylene signaling pathway.

Project description:Plant molecular farming (PMF) is an important growing prospective approach in plant biotechnology; it includes production of recombinant pharmaceutical and industrial proteins in large quantities from engineered plants. Elastin is a major protein component of tissues that require elasticity, it helps keep skin smooth as it stretches to allow normal. Elastin is used as a raw material for the cosmetic industry. In this work, we aimed to use plant as a bioreactor for the expression and production of the full human tropoelastin protein. Agrobacterium- mediated transient expression system into Nicotiana tabacum using syringe agroinfiltration was used to provide fast and convenient way to produce recombinant proteins with greater expression overall the plant leaf. This study aimed to establish an efficient and rapid system for transiently expression and production of human recombinant tropoelastin protein in transgenic N. tabacum plants. Modified elastin (ELN) gene was biosynthesized and cloned into pCambia1390 vector to be used into N. tabacum agroinfilteration. Optimization of codon usage for the human tropoelastin gene, without changing the primary structure of the protein was carried out to ensure high expression in tobacco plants. The obtained data proved that the 5(th) day post-infiltration is the optimum interval to obtain the maximum production of our recombinant protein. Southern blot analysis was able to detect 2175 bp fragment length representing the ELN orf (open reding frame). On the other hand, ELN -expression within plant's tissue was visualized by RT-PCR during the period 3-10 days post agroinfiltration. At the protein level, western and ELISA confirmed the expression of recombinant tropoelastin protein. Western blot analysis detected the tropoelastin protein as parent band at ?70 kDa from freshly extracted protein, while two degraded bands of ?55 and ?45 kDa, representing a pattern of tropoelastin were appeared with frozen samples. This study showed that biosynthetic ELN gene was successfully expressed into N. tabacum leaves using agroinfiltration technique.

Dataset Information

Nuclear-encoded chloroplast ribosomal protein L12 of Nicotiana tabacum: characterization of mature protein and isolation and sequence analysis of cDNA clones encoding its cytoplasmic precursor.

Publications

Nuclear-encoded chloroplast ribosomal protein L12 of Nicotiana tabacum: characterization of mature protein and isolation and sequence analysis of cDNA clones encoding its cytoplasmic precursor.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets