Cloning and Identification of new B-hordein genes
Twenty complete B-hordein gene sequences were cloned from 18 barley materials, designated as BBH-28 to BBH-47, and registered in GenBank (Accession No. ON918636~ON918655). Among them, two each gene sequences were obtained from Yan08221 and 0839-1, while one each sequence was obtained from the other varieties (Table1 and Supplementary file 1). Alignment analyses indicated that the base similarities between the 20 gene sequences were 87.5-99.9%, and no identical sequence were found. The highest similarity was detected between BBH-30 and BBH-34, which had only 1–bp Indel variation at the very beginning of signal peptide region (same in their coding regions). BBH-30, BBH-37, BBH-44, and BBH-47 had relatively low similarities with other gene sequences, all lower than 90%. The results indicated significant sequence variation in B-hordein genes among cultivars used in this study.
Table 1 Fragment length, coding region, and deduced AA number of the 20 cloned genes
Sequence
Number
|
Whole length. of genes
|
Locus of start and stop for coding region
|
Bp No. of coding region
|
No. of Deduced AA
|
(%) Bp Similarity
with others
|
GenBank
Accession No.
|
From variety
|
BBH-28
|
1303
|
268-1215
|
948
|
316
|
87.7-99.6
|
ON918636
|
Supi3
|
BBH-29
|
1301
|
266-1213
|
948
|
316
|
87.5-99.6
|
ON918637
|
Supi4
|
BBH-30
|
1265
|
266-1177
|
912
|
304
|
87.6-100
|
ON918638
|
SuB1403
|
BBH-31
|
1287
|
267-1199
|
933
|
311
|
87.5-99.3
|
ON918639
|
Yan04189
|
BBH-32
|
1302
|
267-1214
|
948
|
316
|
87.5-99.6
|
ON918640
|
Yan06148
|
BBH-33
|
1240
|
267-1139
|
873
|
291
|
87.4-97.5
|
ON918641
|
Yan07216
|
BBH-34
|
1266
|
267-1178
|
912
|
304
|
87.6-100
|
ON918642
|
Yan08221
|
BBH-35
|
1266
|
267-797
|
531
|
pseudogene
|
87.6-99.8
|
ON918643
|
Yan08221
|
BBH-36
|
1153
|
268-1065
|
798
|
266
|
89.0-97.7
|
ON918644
|
Zhu12033
|
BBH-37
|
1236
|
267-1160
|
894
|
298
|
87.1-97.5
|
ON918645
|
Edamai538
|
BBH-38
|
1267
|
267-872
|
606
|
pseudogene
|
87.3-99.8
|
ON918646
|
Longdamai9
|
BBH-39
|
1273
|
267-1199
|
933
|
311
|
87.1-99.6
|
ON918647
|
Gankenpi438
|
BBH-40
|
1266
|
267-1178
|
912
|
304
|
87.4-99.8
|
ON918648
|
0839-1
|
BBH-41
|
1266
|
267-1178
|
912
|
304
|
87.4-99.8
|
ON918649
|
0839-1
|
BBH-42
|
1266
|
267-1178
|
912
|
304
|
87.3-99.8
|
ON918650
|
207-15
|
BBH-43
|
1286
|
267-1163
|
897
|
pseudogene
|
87.2-99.6
|
ON918651
|
WDM2528
|
BBH-44
|
1226
|
266-679
|
414
|
pseudogene
|
88.3-99.3
|
ON918652
|
WDM2563
|
BBH-45
|
1288
|
268-1200
|
933
|
311
|
87.7-98.3
|
ON918653
|
WDM3060
|
BBH-46
|
1286
|
267-737
|
471
|
pseudogene
|
87.2-99.5
|
ON918654
|
WDM2973
|
BBH-47
|
1226
|
266-679
|
414
|
pseudogene
|
88.0-99.3
|
ON918655
|
WDM2975
|
Comparison of new cloned genes with B-hordein genes from GenBank
In total, 72 B-hordein gene sequences with complete coding sequences were obtained by searching the genes against GenBank, including 27 B-hordein genes identified previously by us (Sequence number BBH-1~BBH-27, Accessions ON856174~ON856200). And then, the 20 new cloned genes were compared with above all sequences by means of DNAman. The result showed that their base similarities were 82.9-99.8% (Table 2). It was verified that all the 20 B-hordein sequences were new genes or new allelic variations. The sequence variations among the B-hordein genes were very rich, including SNPs, single base Indels, 3-bp Indels and long-fragment Indels (Table 2 and Supplementary file 1).
Characterization of B-hordein genes and their deduced AA sequences
All the 20 newly-cloned B-hordein genes contained the complete coding sequences, partial promoter sequence (including CATC box and Start codon), and termination sequence (including stop codon and AATAAA polyadenylation signal). The deduced AA sequences showed that 14 genes contained a single open reading frame (ORF) of 798-948bp, encoding 266-316 AA , with a molecular weight of 33.0-36.0KD, and six other sequences were pseudogenes, which have 1-2 stop codons in their coding regions (See Supplementary file 1). Each of the 14 putative functional genes encodes a highly conserved signal peptide of 20-AA residues, directly followed by an N-terminal repetitive domain (Domain Ⅱ) and then a C-terminal conserved domain that can be further divided into three distinctive regions, including a cysteine-rich region (Domain III), a glutamine-rich region (Domain IV), and a C-terminal conserved region (Domain V) (Fig. 1). All the B-hordeins showed a typical set of 8 conserved cysteine (Cys) residues except BBH-42 from a wild barley 207-15, which contained 7 cysteines, with the last one replaced by arginine at the 283 position. Notably, the B-hordeins in cultivars SuB1702 and WDM2566 also contained 7 cysteine residues[13] .
Phylogenetic analysis of B-hordein genes
In order to understand the evolutionary history of barley prolamin genes, phylogenetic analysis was conducted using the 92 complete B-hordein gene sequences available in GenBank and this study. The result showed that the phylogenetic tree was very complicated, and had multilevel branches because of the large numbers and high base similarity of B-hordein genes (81.9-100%). However, they could be distinguished clearly and clustered seperately based on their terminal amino acids of deduced peptides. For example, all the genes and pseudogenes ended with the last AA “Y” of deduced peptides (AY695367, etc,) were classified into one group or two subgroups, while all the genes and pseudogenes terminated with the last AA of “H”(GQ359104, etc.) were also classified into one group or two subgroups except GQ347974 that belonged to a single group (Table 2 and Fig. 2). Other groups included merely the genes and pseudogenes finished with the last AA of “V”.
Table 2. Comparison of the 20 cloned genes with the B-hordein genes from GenBank (%)
Accession
|
BBH-28
|
-29
|
-30
|
-31
|
-32
|
-33
|
-34
|
-35
|
-36
|
-37
|
-38
|
-39
|
-40
|
-41
|
-42
|
-43
|
-44
|
-45
|
-46
|
-47
|
T/S
|
BBH-1
|
95.0
|
94.9
|
99.8
|
99.1
|
94.9
|
87.8
|
99.8
|
99.7
|
97.6
|
87.4
|
99.6
|
98.5
|
99.7
|
99.7
|
99.6
|
98.5
|
88.5
|
98.1
|
98.4
|
88.2
|
IDTRVGV
|
BBH-2
|
94.6
|
94.5
|
98.7
|
98.3
|
94.6
|
88.0
|
98.7
|
98.5
|
97.5
|
87.9
|
98.4
|
99.6
|
98.5
|
98.5
|
98.4
|
99.7
|
88.8
|
97.2
|
99.6
|
88.5
|
IDTRVGV
|
BBH-3
|
94.7
|
94.6
|
98.5
|
98.2
|
94.6
|
87.9
|
98.5
|
98.3
|
97.5
|
87.8
|
98.3
|
99.4
|
98.3
|
98.3
|
98.3
|
99.5
|
88.7
|
97.0
|
99.5
|
88.4
|
V-Pg
|
BBH-4
|
99.2
|
99.2
|
94.9
|
94.6
|
99.2
|
88.2
|
94.9
|
94.7
|
95.6
|
87.9
|
94.6
|
94.3
|
94.7
|
94.7
|
94.6
|
94.3
|
88.6
|
95.2
|
94.3
|
88.1
|
IDTRVGV
|
BBH-5
|
99.6
|
99.5
|
95.1
|
94.9
|
99.5
|
88.6
|
95.1
|
94.9
|
96.2
|
88.6
|
94.9
|
94.5
|
94.9
|
94.9
|
94.9
|
94.6
|
88.9
|
95.4
|
94.6
|
88.6
|
IDTRVGV
|
BBH-6
|
94.9
|
94.9
|
99.8
|
99.1
|
94.9
|
88.5
|
99.8
|
99.6
|
97.4
|
88.8
|
99.5
|
98.4
|
99.6
|
99.6
|
99.5
|
98.4
|
88.8
|
98.0
|
98.3
|
88.7
|
IDTRVGV
|
BBH-7
|
99.4
|
99.3
|
94.9
|
94.6
|
99.3
|
88.7
|
94.9
|
94.7
|
96.0
|
88.7
|
94.6
|
94.3
|
94.7
|
94.7
|
94.6
|
94.3
|
89.0
|
95.2
|
94.3
|
88.6
|
IDTRVGV
|
BBH-8
|
94.4
|
94.3
|
98.5
|
98.3
|
94.3
|
88.0
|
98.5
|
98.3
|
97.4
|
88.3
|
98.2
|
99.4
|
98.3
|
98.3
|
98.2
|
99.5
|
88.5
|
97.0
|
99.4
|
88.2
|
IDTRVGV
|
BBH-9
|
94.9
|
94.9
|
99.8
|
99.1
|
94.9
|
88.4
|
99.8
|
99.6
|
97.5
|
88.4
|
99.5
|
98.4
|
99.6
|
99.6
|
99.5
|
98.4
|
89.0
|
98.0
|
98.3
|
88.8
|
IDTRVGV
|
BBH-10
|
99.5
|
99.5
|
95.1
|
94.9
|
99.5
|
88.7
|
95.1
|
94.9
|
95.9
|
88.2
|
94.9
|
94.5
|
94.9
|
94.9
|
94.9
|
94.6
|
89.0
|
95.4
|
94.6
|
88.6
|
IDTRVGV
|
BBH-11
|
88.9
|
88.8
|
88.9
|
88.7
|
88.8
|
99.5
|
88.9
|
88.9
|
89.5
|
97.1
|
88.6
|
88.2
|
88.7
|
88.7
|
88.6
|
88.3
|
92.8
|
88.8
|
88.4
|
92.4
|
DFWH
|
BBH-12
|
94.5
|
94.4
|
98.5
|
98.1
|
94.4
|
88.1
|
98.5
|
98.3
|
97.3
|
88.1
|
98.3
|
99.5
|
98.3
|
98.3
|
98.3
|
99.5
|
88.8
|
97.0
|
99.5
|
88.5
|
IDTRVGV
|
BBH-13
|
98.6
|
98.5
|
95.1
|
94.8
|
98.5
|
88.6
|
95.1
|
94.9
|
96.0
|
88.3
|
94.9
|
94.3
|
94.9
|
94.9
|
94.9
|
94.3
|
89.3
|
95.3
|
94.4
|
89.0
|
IDTRVGV
|
BBH-14
|
88.5
|
88.4
|
88.1
|
87.9
|
88.4
|
99.6
|
88.1
|
88.1
|
89.2
|
97.1
|
87.9
|
87.5
|
88.0
|
88.0
|
87.9
|
87.6
|
92.6
|
88.0
|
87.7
|
92.2
|
DFWH
|
BBH-15
|
94.5
|
94.4
|
98.5
|
98.1
|
94.4
|
87.9
|
98.5
|
98.3
|
97.2
|
88.1
|
98.3
|
98.7
|
98.3
|
98.3
|
98.3
|
98.8
|
88.8
|
97.0
|
98.7
|
88.5
|
IDTRVGV
|
BBH-16
|
94.3
|
94.2
|
98.5
|
98.1
|
94.2
|
88.0
|
98.5
|
98.3
|
97.3
|
87.9
|
98.3
|
99.5
|
98.3
|
98.3
|
98.3
|
99.5
|
88.8
|
97.2
|
99.5
|
88.5
|
IDTRVGV
|
BBH-17
|
96.1
|
96.0
|
94.2
|
94.1
|
96.1
|
88.0
|
94.2
|
94.1
|
95.1
|
87.8
|
94.0
|
93.8
|
94.1
|
94.1
|
94.0
|
93.8
|
88.6
|
94.5
|
93.8
|
88.3
|
V-Pg
|
BBH-18
|
95.1
|
95.0
|
99.2
|
99.0
|
95.0
|
88.2
|
99.2
|
99.1
|
97.8
|
88.1
|
99.0
|
98.3
|
99.1
|
99.1
|
99.0
|
98.3
|
88.9
|
97.9
|
98.3
|
88.8
|
IDTRVGV
|
BBH-19
|
95.1
|
95.0
|
99.2
|
99.0
|
95.0
|
88.2
|
99.2
|
99.1
|
97.7
|
88.1
|
99.0
|
98.3
|
99.1
|
99.1
|
99.0
|
98.3
|
88.9
|
97.9
|
98.3
|
88.8
|
IDTRVGV
|
BBH-20
|
96.4
|
96.3
|
94.7
|
94.5
|
96.4
|
88.0
|
94.7
|
94.5
|
95.4
|
87.8
|
94.5
|
95.2
|
94.5
|
94.5
|
94.5
|
94.3
|
88.9
|
94.7
|
94.2
|
88.6
|
V-Pg
|
BBH-21
|
96.8
|
96.7
|
95.4
|
95.0
|
96.8
|
89.5
|
95.4
|
95.2
|
96.0
|
89.5
|
95.1
|
94.7
|
95.2
|
95.2
|
95.1
|
94.8
|
90.6
|
95.4
|
94.6
|
90.3
|
IDTRVGV
|
BBH-22
|
99.5
|
99.5
|
95.0
|
94.6
|
99.5
|
88.6
|
95.0
|
95.0
|
95.9
|
88.3
|
94.8
|
94.3
|
94.8
|
94.8
|
94.8
|
94.3
|
89.2
|
95.3
|
94.3
|
88.8
|
V-Pg
|
BBH-23
|
94.6
|
94.5
|
98.7
|
98.3
|
94.6
|
88.3
|
98.7
|
98.5
|
97.1
|
88.5
|
98.4
|
98.8
|
98.5
|
98.5
|
98.4
|
98.9
|
89.1
|
97.2
|
98.8
|
88.8
|
IDTRVGV
|
BBH-24
|
96.2
|
96.1
|
97.4
|
97.0
|
96.1
|
89.0
|
97.4
|
97.2
|
99.5
|
88.8
|
97.2
|
97.2
|
97.2
|
97.2
|
97.2
|
97.1
|
89.4
|
98.1
|
97.0
|
89.1
|
IDTRVGV
|
BBH-25
|
95.0
|
94,9
|
99.1
|
98.9
|
94,9
|
88.5
|
99.1
|
99.0
|
97.7
|
88.6
|
98.9
|
98.2
|
99.0
|
99.0
|
99.1
|
98.3
|
88.9
|
97.9
|
98.2
|
88.8
|
IDTRVGV
|
BBH-26
|
95.5
|
95.4
|
98.3
|
97.7
|
95.4
|
88.4
|
98.3
|
98.1
|
98.1
|
88.7
|
98.0
|
97.1
|
98.1
|
98.1
|
98.0
|
97.2
|
88.9
|
99.5
|
97.1
|
88.7
|
IDTRVGV
|
BBH-27
|
87.8
|
87.7
|
88.1
|
87.9
|
87.7
|
96.5
|
88.1
|
88.1
|
89.0
|
97.9
|
87.9
|
87.4
|
87.9
|
87.9
|
87.9
|
87.7
|
92.3
|
88.0
|
87.6
|
91.8
|
V-Pg
|
AY695367
|
83.1
|
83.3
|
84.0
|
84.0
|
83.3
|
82.9
|
84.0
|
84.0
|
84.2
|
84.3
|
83.8
|
84.1
|
83.8
|
84.0
|
83.8
|
83.9
|
85.3
|
83.4
|
83.8
|
85.3
|
GVGAY
|
AY695368
|
83.5
|
83.6
|
84.3
|
84.1
|
83.6
|
83.3
|
84.3
|
84.3
|
84.6
|
84.7
|
84.0
|
84.3
|
84.0
|
84.3
|
84.0
|
84.0
|
85.7
|
83.5
|
83.9
|
85.7
|
GVGAY
|
AY695369
|
83.8
|
84.0
|
84.5
|
84.5
|
84.0
|
83.5
|
84.5
|
84.5
|
84.7
|
84.9
|
84.3
|
84.8
|
84.3
|
84.5
|
84.3
|
84.6
|
86.3
|
83.8
|
84.5
|
86.0
|
GVGGY
|
AY700801
|
85.9
|
85.8
|
86.2
|
86.1
|
85.6
|
87.6
|
86.2
|
86.2
|
86.3
|
88.7
|
86.0
|
86.0
|
86.0
|
86.2
|
86.0
|
86.2
|
88.5
|
86.1
|
86.2
|
88.2
|
GVGPY
|
AY700802
|
86.4
|
86.3
|
87.1
|
87.0
|
86.2
|
88.0
|
87.1
|
87.1
|
87.2
|
89.3
|
86.8
|
86.9
|
86.8
|
87.1
|
86.8
|
87.1
|
88.9
|
87.0
|
87.1
|
88.6
|
GVGPY
|
AY700803
|
86.5
|
86.4
|
87.2
|
87.1
|
86.3
|
88.1
|
87.2
|
87.2
|
87.3
|
89.4
|
87.0
|
87.0
|
87.0
|
87.2
|
87.0
|
87.2
|
89.0
|
87.1
|
87.2
|
88.6
|
GVGPY
|
AY700804
|
87.0
|
86.8
|
87.6
|
87.5
|
86.7
|
87.8
|
87.6
|
87.6
|
87.7
|
89.0
|
87.4
|
87.4
|
87.4
|
87.6
|
87.4
|
87.6
|
88.5
|
87.5
|
87.6
|
88.1
|
GVGPY
|
AY700805
|
86.4
|
86.3
|
86.8
|
86.7
|
86.2
|
87.4
|
86.8
|
86.8
|
86.9
|
88.4
|
86.6
|
86.6
|
86.6
|
86.8
|
86.6
|
86.8
|
88.1
|
86.7
|
86.8
|
87.8
|
GVGPY
|
AY700806
|
86.6
|
86.4
|
86.9
|
86.8
|
86.3
|
87.5
|
86.9
|
86.9
|
87.0
|
88.5
|
86.7
|
86.7
|
86.7
|
86.9
|
86.7
|
86.9
|
88.2
|
86.8
|
86.9
|
87.9
|
GVGPY
|
AY700807
|
85.9
|
85.8
|
86.2
|
86.1
|
85.6
|
87.6
|
86.2
|
86.2
|
86.3
|
88.7
|
86.0
|
86.0
|
86.0
|
86.2
|
86.0
|
86.2
|
88.5
|
86.1
|
86.2
|
88.2
|
Y-Pg
|
DQ148297
|
89.7
|
89.6
|
89.3
|
88.8
|
89.6
|
94.9
|
89.3
|
89.1
|
90.0
|
95.9
|
89.1
|
88.5
|
89.1
|
89.1
|
89.1
|
88.8
|
91.6
|
89.2
|
88.7
|
91.2
|
IDTRVGV
|
DQ178602
|
88.6
|
88.5
|
88.5
|
88.4
|
88.5
|
96.6
|
88.5
|
88.5
|
89.3
|
95.4
|
88.2
|
87.8
|
88.3
|
88.5
|
88.2
|
87.9
|
93.3
|
88.2
|
88.0
|
92.0
|
DFWH
|
DQ189997
|
88.2
|
88.1
|
88.1
|
87.9
|
88.2
|
99.4
|
88.1
|
88.1
|
89.2
|
96.6
|
87.9
|
87.4
|
88.0
|
88.0
|
87.9
|
87.6
|
92.1
|
88.0
|
87.7
|
91.7
|
H-Pg
|
DQ267476
|
88.2
|
88.1
|
88.0
|
87.8
|
88.1
|
99.1
|
88.0
|
88.0
|
89.1
|
96.5
|
87.8
|
87.4
|
87.9
|
87.9
|
87.8
|
87.5
|
92.0
|
87.9
|
87.6
|
91.6
|
H-Pg
|
DQ267477
|
88.2
|
88.1
|
88.1
|
87.9
|
88.2
|
99.2
|
88.1
|
88.1
|
89.1
|
96.6
|
87.9
|
87.4
|
88.0
|
88.0
|
87.9
|
87.6
|
92.1
|
88.0
|
87.7
|
91.7
|
H-Pg
|
DQ267478
|
94.7
|
94.5
|
99.7
|
98.9
|
94.5
|
88.5
|
99.7
|
99.5
|
97.2
|
88.7
|
99.4
|
98.4
|
99.5
|
99.5
|
99.5
|
98.4
|
89.6
|
97.7
|
98.4
|
89.4
|
IDTRVGV
|
DQ267479
|
88.9
|
88.8
|
88.9
|
88.9
|
88.8
|
98.7
|
88.9
|
88.9
|
89.6
|
96.9
|
88.7
|
88.5
|
88.7
|
88.7
|
88.7
|
88.5
|
92.7
|
88.8
|
88.7
|
92.3
|
DFWH
|
DQ267480
|
87.9
|
87.8
|
87.9
|
87.8
|
87.8
|
98.4
|
88.0
|
88.0
|
89.1
|
96.3
|
87.7
|
87.5
|
87.8
|
87.8
|
87.7
|
87.6
|
92.1
|
87.8
|
87.7
|
91.7
|
H-Pg
|
DQ267481
|
88.0
|
87.9
|
87.9
|
87.7
|
87.9
|
98.4
|
87.9
|
87.9
|
89.2
|
96.1
|
87.6
|
87.4
|
87.7
|
87.7
|
87.6
|
87.5
|
91.9
|
87.8
|
87.8
|
91.5
|
H-Pg
|
DQ826387
|
90.0
|
89.8
|
91.7
|
91.5
|
89.8
|
92.6
|
91.7
|
91.7
|
91.8
|
92.5
|
91.5
|
90.9
|
91.6
|
91.6
|
91.5
|
90.9
|
92.5
|
91.4
|
91.0
|
92.1
|
VGPSVGV
|
GQ342970
|
95.4
|
95.2
|
99.2
|
98.5
|
95.2
|
88.5
|
99.2
|
99.0
|
98.0
|
88.8
|
98.9
|
98.1
|
99.0
|
99.0
|
98.9
|
98.1
|
89.5
|
98.5
|
98.1
|
89.2
|
IDTRVGV
|
GQ342971
|
94.7
|
94.5
|
99.6
|
98.9
|
94.5
|
88.4
|
99.6
|
99.4
|
97.1
|
88.6
|
99.3
|
98.3
|
99.4
|
99.4
|
99.4
|
98.3
|
89.3
|
97.7
|
98.3
|
89.0
|
IDTRVGV
|
GQ342972
|
95.1
|
95.0
|
98.9
|
98.3
|
95.0
|
88.5
|
98.9
|
98.8
|
97.9
|
88.8
|
98.7
|
97.8
|
98.8
|
98.8
|
98.7
|
97.8
|
89.2
|
98.4
|
97.8
|
88.9
|
IDTRVGV
|
GQ342973
|
97.0
|
96.8
|
96.9
|
96.4
|
96.9
|
87.4
|
96.9
|
96.7
|
94.9
|
87.4
|
96.6
|
95.8
|
96.7
|
96.7
|
96.7
|
95.7
|
88.3
|
95.5
|
95.7
|
87.9
|
IDTRVGV
|
GQ342974
|
93.7
|
93.5
|
97.5
|
97.1
|
93.5
|
90.5
|
97.5
|
97.5
|
95.2
|
89.7
|
97.2
|
96.6
|
97.3
|
97.3
|
97.3
|
96.4
|
89.5
|
95.8
|
96.5
|
89.3
|
DFWH
|
GQ342975
|
98.4
|
98.3
|
96.3
|
95.7
|
98.3
|
87.9
|
96.3
|
96.1
|
95.5
|
87.8
|
96.0
|
95.3
|
96.1
|
96.1
|
96.1
|
95.3
|
88.7
|
94.9
|
95.3
|
88.3
|
IDTRVGV
|
GQ342976
|
94.5
|
94.3
|
99.5
|
98.7
|
94.3
|
88.4
|
99.5
|
99.3
|
97.0
|
88.7
|
99.2
|
98.2
|
99.3
|
99.3
|
99.3
|
98.3
|
89.6
|
97.6
|
98.3
|
89.4
|
IDTRVGV
|
GQ342977
|
87.8
|
87.7
|
87.7
|
87.7
|
87.7
|
97.8
|
87.7
|
87.7
|
89.1
|
95.5
|
87.5
|
87.4
|
87.5
|
87.5
|
87.5
|
87.5
|
92.5
|
87.8
|
87.7
|
92.1
|
H-Pg
|
GQ359102
|
88.9
|
88.8
|
88.7
|
88.7
|
88.8
|
99.4
|
88.7
|
88.7
|
89.4
|
96.9
|
88.7
|
88.5
|
88.6
|
88.6
|
88.5
|
88.5
|
92.7
|
88.8
|
88.7
|
92.3
|
H-Pg
|
GQ359103
|
89.2
|
89.1
|
88.8
|
88.8
|
89.1
|
97.4
|
88.8
|
88.8
|
89.5
|
95.8
|
88.5
|
88.4
|
88.6
|
88.8
|
88.5
|
88.5
|
94.4
|
88.8
|
88.6
|
94.2
|
H-Pg
|
GQ359104
|
87.7
|
87.6
|
87.5
|
87.5
|
87.6
|
98.1
|
87.6
|
87.6
|
89.0
|
96.0
|
87.3
|
87.3
|
87.4
|
87.4
|
87.3
|
87.3
|
91.7
|
87.6
|
87.5
|
91.3
|
H-Pg
|
GQ359105
|
88.5
|
88.4
|
88.4
|
88.2
|
88.4
|
99.3
|
88.5
|
88.5
|
89.1
|
96.6
|
88.2
|
88.0
|
88.3
|
88.3
|
88.2
|
88.1
|
92.1
|
88.4
|
88.2
|
91.7
|
H-Pg
|
GQ359106
|
88.2
|
88.1
|
88.2
|
88.0
|
88.2
|
99.0
|
88.2
|
88.2
|
88.8
|
96.4
|
88.0
|
87.8
|
88.1
|
88.1
|
88.0
|
87.8
|
91.8
|
88.2
|
88.0
|
91.4
|
H-Pg
|
GQ359107
|
88.4
|
88.3
|
88.4
|
88.2
|
88.3
|
99.2
|
88.4
|
88.4
|
89.0
|
96.5
|
88.1
|
87.9
|
88.2
|
88.2
|
88.1
|
88.0
|
92.0
|
88.3
|
88.1
|
91.6
|
H-Pg
|
KP866149
|
88.4
|
88.3
|
88.5
|
88.2
|
88.3
|
96.3
|
88.5
|
88.5
|
89.0
|
97.4
|
88.3
|
88.1
|
88.4
|
88.4
|
88.3
|
88.1
|
91.8
|
88.3
|
88.2
|
91.4
|
VGTRVGV
|
KP872301
|
90.8
|
90.8
|
90.8
|
90.6
|
90.8
|
95.2
|
90.8
|
90.8
|
91.6
|
95.8
|
90.6
|
90.3
|
90.6
|
90.8
|
90.6
|
90.2
|
95.7
|
90.6
|
90.3
|
95.4
|
VGPXVGV
|
KP872302
|
87.6
|
87.6
|
88.2
|
88.3
|
87.5
|
89.8
|
88.2
|
88.2
|
88.3
|
89.4
|
87.9
|
88.1
|
88.0
|
88.1
|
88.0
|
88.2
|
89.1
|
87.9
|
88.2
|
88.8
|
VGTGVGV
|
KP872303
|
87.6
|
87.6
|
87.8
|
87.6
|
87.5
|
96.7
|
87.8
|
87.8
|
88.7
|
98.2
|
87.6
|
87.3
|
87.6
|
87.6
|
87.6
|
87.4
|
91.3
|
87.8
|
87.3
|
91.0
|
VGTRVGV
|
KP872304
|
88.2
|
88.2
|
88.3
|
88.1
|
88.1
|
97.5
|
88.3
|
88.3
|
89.1
|
98.7
|
88.1
|
87.8
|
88.2
|
88.2
|
88.1
|
88.0
|
91.6
|
88.1
|
87.9
|
91.3
|
VGTRVGV
|
KP872305
|
89.2
|
89.2
|
89.1
|
88.9
|
89.2
|
98.9
|
89.1
|
89.1
|
90.1
|
99.0
|
88.9
|
88.5
|
88.9
|
88.9
|
88.9
|
88.5
|
92.2
|
89.0
|
88.6
|
92.0
|
VGTRVXV
|
KP872306
|
88.6
|
88.6
|
88.7
|
88.4
|
88.5
|
97.3
|
88.7
|
88.7
|
89.2
|
98.0
|
88.4
|
88.1
|
88.5
|
88.5
|
88.4
|
88.1
|
91.9
|
88.5
|
88.2
|
91.6
|
VGTRVGV
|
X03103
|
89.2
|
89.1
|
88.8
|
88.5
|
89.2
|
93.9
|
88.8
|
88.8
|
89.4
|
93.2
|
88.5
|
88.1
|
88.6
|
88.8
|
88.5
|
88.2
|
96.2
|
88.5
|
88.4
|
95.9
|
VGPSVGV
|
X53690
|
88.1
|
88.0
|
88.0
|
87.7
|
88.0
|
99.8
|
88.0
|
88.0
|
88.8
|
95.9
|
87.8
|
87.5
|
87.9
|
87.9
|
87.8
|
87.5
|
92.5
|
87.9
|
87.7
|
92.1
|
DFWH
|
X53691
|
88.6
|
88.5
|
88.4
|
88.2
|
88.6
|
96.1
|
88.4
|
88.4
|
89.3
|
94.6
|
88.2
|
87.9
|
88.3
|
88.3
|
88.2
|
87.9
|
94.8
|
88.3
|
88.0
|
94.4
|
V-Pg
|
X87232
|
87.0
|
86.9
|
86.5
|
86.6
|
86.9
|
95.0
|
86.6
|
86.6
|
87.4
|
97.6
|
86.3
|
86.1
|
86.4
|
86.4
|
86.3
|
86.2
|
90.9
|
86.8
|
86.4
|
90.5
|
VGTRVGV
|
T/S
|
IDTR
-VGV
|
IDTR
-VGV
|
IDTR
-VGV
|
IDTR
-VGV
|
IDTR
-VGV
|
DFWH
|
IDTR
-VGV
|
V-Pg
|
IDTR
-VGV
|
VGTR
-VGV
|
V-Pg
|
IDTR
-VGV
|
IDTR
-VGV
|
IDTR
-VGV
|
IDTR
-VGV
|
V-Pg
|
V-Pg
|
IDTR
-VGV
|
V-Pg
|
V-Pg
|
T/S
|
Note:①“T/S” means “Terminal sequences” of the deduced peptides; ②“V-Pg, H-Pg and Y-Pg” means “V-type, H-type and Y-type pseudogene”.