Background
Methods
Samples
Sequencing of the full length RDL coding sequence from Anopheles sinensis
PCR amplification of AsRDL fragments
Haplotype reconstruction and confirmation
Genealogical analysis
Results
Identification of AsRDL gene
Identification of naturally occurring genetic mutations in AsRDL
296A | 296S | 327V | 327I | 345T | 345S | 349M | 349L | 356R | 356G | 357K | 357L | 357M | 357N | 360F | 360L | 408Q | 408H | 473G | 473S | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
NN | 1.000 | 0.636 | 0.364 | 0.591 | 0.409 | 0.985 | 0.015 | 1.000 | 0.924 | 0.076 | 0.985 | 0.015 | 1.000 | 1.000 | ||||||
YL | 1.000 | 0.650 | 0.350 | 0.583 | 0.417 | 0.950 | 0.050 | 1.000 | 0.967 | 0.033 | 0.983 | 0.017 | 1.000 | 0.983 | 0.017 | |||||
HZ | 1.000 | 0.882 | 0.118 | 0.529 | 0.471 | 1.000 | 1.000 | 0.882 | 0.088 | 0.030 | 1.000 | 1.000 | 0.706 | 0.294 | ||||||
BS | 0.182 | 0.818 | 0.795 | 0.205 | 0.648 | 0.352 | 1.000 | 1.000 | 0.966 | 0.011 | 0.023 | 0.989 | 0.011 | 0.989 | 0.011 | 1.000 | ||||
WZ | 1.000 | 0.738 | 0.262 | 0.548 | 0.452 | 0.952 | 0.048 | 0.976 | 0.024 | 0.976 | 0.024 | 1.000 | 1.000 | 0.881 | 0.119 | |||||
LZ | 0.229 | 0.771 | 0.917 | 0.083 | 0.625 | 0.375 | 0.979 | 0.021 | 1.000 | 1.000 | 1.000 | 1.000 | 0.792 | 0.208 | ||||||
GL | 0.093 | 0.907 | 0.704 | 0.296 | 0.741 | 0.259 | 0.944 | 0.056 | 1.000 | 0.907 | 0.093 | 1.000 | 1.000 | 0.870 | 0.130 | |||||
HC | 1.000 | 0.397 | 0.603 | 0.776 | 0.224 | 0.983 | 0.017 | 1.000 | 0.983 | 0.017 | 1.000 | 1.000 | 1.000 | |||||||
GG | 1.000 | 0.733 | 0.267 | 0.700 | 0.300 | 1.000 | 1.000 | 0.933 | 0.067 | 1.000 | 1.000 | 0.833 | 0.167 |
Distribution and frequency of resistance-related mutations
Residue sites | Populations | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
296 | 327 | 345 | NN | YL | HZ | BS | WZ | LZ | GL | HC | GG | |
Combination 1 |
S/S
|
I/I
| T/T | 12.1 | 13.3 | 2.3 | 4.2 | 3.7 | 34.5 | 6.7 | ||
Combination 2 |
S/S
|
I/V |
S/S
| 2.3 | 3.4 | |||||||
Combination 3 |
S/S
|
I/V | T/S
| 36.4 | 23.3 | 23.5 | 11.4 | 33.3 | 4.2 | 22.2 | 24.1 | 26.7 |
Combination 4 |
S/S
|
I/V | T/T | 12.1 | 20.0 | 18.2 | 19.0 | 25.9 | 24.1 | 13.3 | ||
Combination 5 |
S/S
| V/V |
S/S
| 15.2 | 23.3 | 23.5 | 9.1 | 19.0 | 16.7 | 3.7 | 3.4 | |
Combination 6 |
S/S
| V/V | T/S
| 15.2 | 13.3 | 23.5 | 25.0 | 19.0 | 29.2 | 18.5 | 6.9 | 33.3 |
Combination 7 |
S/S
| V/V | T/T | 9.1 | 6.7 | 29.4 | 2.3 | 9.5 | 12.5 | 7.4 | 3.4 | 20.0 |
Combination 8 |
S/A | I/V | T/T | 4.5 | 4.2 | 3.7 | ||||||
Combination 9 |
S/A | V/V | T/S
| 11.4 | 8.3 | 3.7 | ||||||
Combination 10 |
S/A | V/V | T/T | 6.8 | 8.3 | 11.1 | ||||||
Combination 11 | A/A | V/V | T/T | 6.8 | 12.5 |
Haplotype diversity of AsRDL gene
Haplotype | Polymorphic sites | Intron type | 345aa | NN | YL | HZ | BS | WZ | LZ | GL | HC | GG | Total numbers |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
H1 | ACCCTGATTGAACAAGTCGGCCGGGGGT | [1] | T | 4 | 2 | 1 | 4 | 3 | 4 | 3 | 6 | 2 | 29 |
H2
| ACCCTGATTGTACAAGTCGGCCGGGGGT | [1] | S | 7 | 9 | 7 | 2 | 3 | 1 | 29 | |||
H3 | ACCCTGCTTGAACAAGTCGGCCGGGGGT | [2] | T | 26 | 28 | 9 | 22 | 13 | 10 | 21 | 28 | 10 | 167 |
H4 | ACCCTGCTTGAACAAGTCGGCCGGGGGC | [2] | T | 1 | 1 | ||||||||
H5 | ACCCTGCTTGAACAAGTCGGCCGGGGAT | [2] | T | 3 | 1 | 2 | 3 | 2 | 11 | ||||
H6 | ACCCTGCTTGAACAAGTCGGCCGGATGC | [2] | T | 1 | 1 | ||||||||
H7 | ACCCTGCTTGAACAAGTCGGCCCGATGC | [2] | T | 1 | 1 | ||||||||
H8 | ACCCTGCTTGAACAAGTCGGTCGGGGGT | [2] | T | 1 | 1 | ||||||||
H9 | ACCCTGCTTGAACAAGTCGACCGGGGGT | [2] | T | 1 | 1 | ||||||||
H10 | ACCCTGCTTGAACAAGTGGGCCGGGGGT | [2] | T | 1 | 1 | ||||||||
H11 | ACCCTGCTTGAACAAGCCGGCCGGGGGT | [2] | T | 1 | 1 | 1 | 2 | 5 | |||||
H12 | ACCCTGCTTGAACAACTCGGCCGGGGGT | [2] | T | 1 | 1 | ||||||||
H13 | ACCCTGCTTGAACATGTCGGCCGGGGGT | [2] | T | 5 | 2 | 1 | 1 | 1 | 10 | ||||
H14 | ACCCTGCTTGAACTTGTCGGCCGGGGGT | [2] | T | 1 | 1 | ||||||||
H15 | ACCCTGCTTGAAGAAGTCGGCCGGGGGT | [2] | T | 1 | 1 | ||||||||
H16 | ACCCTGCTTGATCAAGTCGGCCGGGGGT | [2] | T | 1 | 1 | 1 | 1 | 4 | |||||
H17
| ACCCTGCTTGTACAAGTCGGCCGGGGGT | [2] | S | 13 | 12 | 2 | 17 | 8 | 2 | 2 | 4 | 3 | 63 |
H18
| ACCCTGCTTGTACAAGTCGGCCGGGGAT | [2] | S | 1 | 7 | 4 | 8 | 4 | 3 | 27 | |||
H19
| ACCCTGCTTGTACAAGTCGGCCGGGTGC | [2] | S | 1 | 1 | ||||||||
H20
| ACCCTGCTTGTACAAGTCGGCCGGATGT | [2] | S | 1 | 1 | 2 | |||||||
H21
| ACCCTGCTTGTACAAGTCGGCCGGATGC | [2] | S | 6 | 1 | 1 | 5 | 1 | 1 | 4 | 6 | 25 | |
H22
| ACCCTGCTTGTTCAAGTCGGCCGGGGGT | [2] | S | 1 | 1 | 2 | |||||||
H23 | ACCCTGCTTAAACAAGTCGGCCGGGGGT | [3] | T | 1 | 2 | 1 | 4 | 8 | |||||
H24 | ACCCTGCTTAAACATGTCGGCCGGGGGT | [3] | T | 1 | 1 | 2 | |||||||
H25 | ACCCCGCACGAACAAGTCGGCCGGGGGT | [4] | T | 2 | 18 | 5 | 1 | 7 | 33 | ||||
H26 | ACCCCGCACGAACAAGTCGGCCGGAGGT | [4] | T | 1 | 1 | ||||||||
H27 | ACCCCGCACGAACAAGTCGGCTGGGGGT | [4] | T | 1 | 1 | ||||||||
H28 | ACCCCGCACGAACAAGTCAGCCGGGGGT | [4] | T | 1 | 1 | ||||||||
H29 | ACCCCGCACGAACATGTCGGCCGGGGGT | [4] | T | 3 | 1 | 1 | 2 | 7 | |||||
H30 | ACCCCGCACGAACATGTCGGCCGAGGGT | [4] | T | 1 | 1 | 2 | |||||||
H31 | ACCCCGCACGATCAAGTCGGCCGGGGGT | [4] | T | 2 | 1 | 3 | 1 | 7 | |||||
H32
| ACCCCGCACGTACAAGTCGGCCGGGGGT | [4] | S | 1 | 2 | 3 | |||||||
H33
| ACCCCGCACGTACAAGTCGGCTGGGGGT | [4] | S | 6 | 2 | 7 | 3 | 2 | 20 | ||||
H34 | ACCTTGCTTGAACAAGTCGGCCGGGGGT | [5] | T | 2 | 1 | 3 | 6 | ||||||
H35 | ACCTTGCTTGAACAAGTCGGCCGAGGGT | [5] | T | 2 | 2 | ||||||||
H36 | ACCTTCCTTGAACAAGTCGGCCGGGGGT | [6] | T | 1 | 1 | ||||||||
H37 | ACACCGCACGAACAAGTCGGCCGGGGGT | [7] | T | 1 | 1 | ||||||||
H38 | TTCCCGCACGAACAAGTCGGCCGGGGGT | [8] | T | 1 | 1 | ||||||||
2N | 66 | 60 | 34 | 88 | 42 | 48 | 54 | 58 | 30 | 480 |