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发布于:2020-8-3 17:50:11  访问:14 次 回复:0 篇
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Genes, see Supplemental Table S1). 1 of these genes is AT
One of these genes is AT2G31040, the Arabidopsis equivalent on the MRK-740 Technical Information Chlamydomonas reinhardtii gene CGLD22 (Merchant et al., 2007), which was later renamed CGL160 (Karpowicz et al., 2011). AtCGL160, like its counterparts from other KZR-616 web vascular plants, mosses, and green algae, consists of four transmembrane (TM)-spanning a-helices (hereafter named the "membrane domain") in its C-terminal region (Fig. Neither complementary DNA (cDNA) from an amplicon located 39 of the insertion (amplicon 1 in Fig. 2A) nor from an amplicon situated 59 of the insertion (amplicon 2) might be obtained from atcgl160-1 plants, whereas both portions of AtCGL160 transcripts had been detectable in atcgl160-2 (Fig. 2B). To analyze AtCGL160 expression in the protein level, the N-terminal portion of AtCGL160 was heterologously expressed in, and purified from, E.Genes, see Supplemental Table S1). A single of those genes is AT2G31040, the Arabidopsis equivalent of your Chlamydomonas reinhardtii gene CGLD22 (Merchant et al., 2007), which was later renamed CGL160 (Karpowicz et al., 2011). Within the following, we designate AT2G31040 as AtCGL160 and its C. reinhardtii ortholog as CrCGL160. AtCGL160 codes to get a protein of 350 amino acids containing an N-terminal chloroplast transit peptide (cTP) of 46 amino acids, as predicted by ChloroP (Emanuelsson et al., 1999), such that the mature protein should have a molecular mass of around 34 kD. AtCGL160, like its counterparts from other vascular plants, mosses, and green algae, includes 4 transmembrane (TM)-spanning a-helices (hereafter named the "membrane domain") in its C-terminal region (Fig. 1A). CGL160 from the moss Physcomitrella patens shows 53 /40 similarity/identity to AtCGL160, while the values for CrCGL160 are lower (38 /25 similarity/identity). Cross-species sequence conservation (Fig. 1A) is more pronounced inside the membrane domain (84 /25.7 consensus positions/identity positions; amino acids 20950 of AtCGL160) than inside the N-terminal segment (69.9 /6.7 ; amino acids 4708 of AtCGL160). Here, "identity positions" refers to positions with identical amino acids in every species, whereas "consensus positions" refers to positions at which no less than 50 of species examined show identical or related residues. The GreenCut consists of genes found in photosynthetic eukaryotes but not in nonphotosynthetic organisms, whether prokaryotes or eukaryotes. Nonetheless, the conserved C-terminal portion of AtCGL160 is distantly associated for the items of bacterial atp1/uncI genes, for example the atp1/sll1321 gene in Synechocystis sp. PCC6803 (Grossman et al., 2010; Karpowicz et al., 2011) and the uncI gene in E. coli (Walker et al., 1984). While sequence alignments in the C-terminal membrane domain of AtCGL160 with the products of atp1/uncI genes from various cyanobacteria and E. coli showed only moderate general similarity, the relative positions of four predicted TM-spanning a-helices coincided (Fig. 1B). This suggests that the secondary structure of this domain is conserved between eukaryotic CGL160 and bacterial Atp1/UncIPlant Physiol. Vol. 165,Two distinctive Arabidopsis transfer DNA (T-DNA) insertion lines for AtCGL160 have been identified within the SIGnAL database (http://signal.salk.edu/cgi-bin/tdnaexpress) and obtained in the Nottingham Arabidopsis Stock Centre. The web-sites in the T-DNA insertions have been determined by PCR amplification and sequencing in the regions flanking the T-DNA borders.
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