Molecular cloning and identification of eukaryotic translation initiation factor 1 family genes (eIF1, eIF1A and eIF1B) in Leymus chinensis (Trin.)

Figures & data

Table 1. Primer names, sequences and annealing temperatures (TMs) for PCR amplifications of the eIF1 family genes.

eIF1 family gene	Primer name	Primer direction	Primer sequence (5′–3′)	TMs (°C)
eIF1	eIF1-F1	Forward	TTC ATG TCT GAT CTT GAC ATT	60
	eIF1-R1	Reverse	TGC CTA AAA TCC GTG AAT CTT
eIF1A	eIF1A-F1	Forward	ATG CCG AAG AAC AAG GGT AAG	53
	eIF1A-R1	Reverse	GAT CTT GTC GAT GTC CTC GTC
eIF1B	eIF1B-F1	Forward	ATG TCC GCT ATC CAG AAC CTC	61
	eIF1B-R1	Reverse	TAG AAC CCG TGG ACC TTG AG

Note: Underlined parts show the translation start codon and end codon in the forward and reverse primer of eIF1, respectively.

Figure 1. DNA sequence alignment of eIF1 from different plant species.

Note: AteIF1 (accession number: BT000649) from A. thaliana, OseIF1 (accession number: AF094774) from O. sativa, TaeIF1 (accession number: AF508969) from T. aestivum and ZmeIF1 (accession number: AF034944) from Z. mays. The two red parts are the newly designed forward and reverse primers’ binding sites, respectively.

Figure 2. DNA sequence alignment of eIF1A from different plant species.

Note: HveIF1A (accession number: AK373449) from H. vulgare, OseIF1A-1 (accession number: AJ563657) and OseIF1A-2 (accession number: AJ563658) from O. sativa, TaeIF1A-1 (accession number: WHTEIF1A) and TaeIF1A-2 (accession number: AK332835) from T. aestivum, and ZmeIF1A (accession number: NM001155525) from Z. mays. The two red parts are the newly designed forward and reverse primer binding sites, respectively.

Figure 3. DNA sequence alignment of eIF1B from different species.

Note: SseIF1B (accession number: BT057513) from S. salar, DreIF1B (accession number: BC067620) from D. rerio, HseIF1B (accession number: BC006996) from H. sapiens and MmeIF1B (accession number: NM026892) from M. musculus. The two red parts are the newly designed forward and reverse primer binding sites, respectively.

Figure 4. Bioinformatics analysis of L. chinensis eIF1 gene and protein.

Note: (A) Nucleotide and deduced amino acid sequence of LceIF1. The amino acid is indicated by the one letter designation below the nucleotide sequence. The putative binding sites are marked as red, boldface letters. The predicted RNA interaction sites are marked as blue, boldface letters. The mutations affecting start sites are underlined. (B) Multiple sequence alignment among EIF1 protein sequences, including AtEIF1 (accession number: AAN18215) from A. thaliana, OsEIF1 (accession number: AAC67556) from O. sativa, TaEIF1 (accession number: AAM34279) from T. aestivum, ZmEIF1 (accession number: AAB88615) from Z. mays, and LcEIF1 (accession number: ADD62702). (C) Phylogenetic tree analysis among these EIF1 protein sequences.

Figure 5. Bioinformatics analysis of L. chinensis eIF1A gene and protein.

Note: (A) Nucleotide and deduced amino acid sequence of LceIF1A. The start codon and ending codon are marked red. The conserved oligonucleotide-binding site is underlined. (B) Multiple sequence alignment among EIF1A protein sequences, including AtEIF1A (accession number: BAK04646) from A. thaliana, OsEIF1A-1 (accession number: CAD91550) and OsEIF1A-2 (accession number: CAD91551) from O. sativa, ZmEIF1A-1 (accession number: NP001148997) and ZmEIF1A-2 (accession number: ACG49138) from Z. mays and LcEIF1A (accession number: ADI47121). (C) Phylogenetic tree analysis among these EIF1A protein sequences.

Figure 6. Bioinformatics analysis of L. chinensis eIF1B gene and protein.

Note: (A) Nucleotide and deduced amino acid sequence of LceIFB. (B) Multiple sequence alignment among EIF1B protein sequences, including SsEIF1B (accession number: ACM09385) from S. salar, DrEIF1B (accession number: AAH67620) from D. rerio, HsEIF1B (accession number: AAH06996) from H. sapiens, MmEIF1B (accession number: NP081168) from M. musculus and LcEIF1B (accession number: ADN77997). (C) Phylogenetic tree analysis among these EIF1B protein sequences.

Table 2. Amino acid analysis of protein EIF1B in different organisms.

Organism	Protein amino acid number (aa)	Accession number	Description
Homo sapiens	133	NP005866	EIF1B
Pan troglodytes	113	XP001135385	EIF1B
Canis lupus familiaris	113	XP534229	EIF1B
Bos taurus	113	NP001032687	EIF1B
Mus musculus	113	NP081168	EIF1B
Rattus norvegicus	113	XP217294	EIF1B
Gallus gallus	113	XP418815	EIF1B
Danio rerio	113	NP956587	zgc:56676
Schizosaccharomyces pombe	109	NP595863	EIF1
Saccharomyces cerevisiae	108	NP014155	Sui1p
Kluyveromyces lactis	108	XP452335	Hypotheticalprotein
Eremothecium gossypii	108	NP985312	AER457Wp
Magnaporthe grisea	115	XP001414994	EIF1/SUI1
Neurospora crassa	169	XP329171	Hypothetical protein
Arabidopsis thaliana	112	NP568818	EIF1/SUI1

Note: Amino acid (aa).