Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 152-152.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0064
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YAO Qiong(), QUAN Lin-fa, XU Shu, DONG Yi-zhi, LI Wen-jing, CHI Yan-yan, CHEN Bing-xu()
Received:
2021-01-15
Online:
2021-10-26
Published:
2021-11-12
Contact:
CHEN Bing-xu
E-mail:joanyao_0603@163.com;gzchenbx@163.com
YAO Qiong, QUAN Lin-fa, XU Shu, DONG Yi-zhi, LI Wen-jing, CHI Yan-yan, CHEN Bing-xu. Gene Cloning of 2 Octopamine Receptors from Eocanthecona furcellata and Effects of Chemical Pesticide on Its Expression[J]. Biotechnology Bulletin, 2021, 37(10): 152-152.
用途Purpose | 基因Gene | 5'扩增引物 primers for 5' RACE | 3'扩增引物 primers for 3' RACE |
---|---|---|---|
巢式PCR Nested gene-specific primers | EfOctβ1R | CAATATGGTCGTACTTGGTAG | AAAGAGTGGAAACAGCTTCT |
AGACTACTGCAATTCGTGA | AACAGCTTCTTGACAAATGG | ||
EfOctβ2R | CGCTCCATGTAGTACAAGT | ACTTGTACTACATGGAGCG | |
CACAGCGATACAGGACAG | AGTGGGCCAAGATTGTACG | ||
基因Gene | 5'扩增引物 5' primers | 3'扩增引物3' primers | |
实时荧光定量PCR RT-qPCR | EfOctβ1R | CCTGTGGGTATTGTCGTTCGTC | CGTAGCCTGCGTCGTTGGTTA |
EfOctβ2R | TATCGTTCTGGGTGCCTGGAG | CGGACATAGATGCTGCGTAAC | |
EfEF1a | CCGCCAACATCACTACTGAAGT | GCAACATAACCACGACGCAAT |
Table 1 Primers for EfOctβ1R and EfOctβ2R cloning and qRT-PCR used in this study
用途Purpose | 基因Gene | 5'扩增引物 primers for 5' RACE | 3'扩增引物 primers for 3' RACE |
---|---|---|---|
巢式PCR Nested gene-specific primers | EfOctβ1R | CAATATGGTCGTACTTGGTAG | AAAGAGTGGAAACAGCTTCT |
AGACTACTGCAATTCGTGA | AACAGCTTCTTGACAAATGG | ||
EfOctβ2R | CGCTCCATGTAGTACAAGT | ACTTGTACTACATGGAGCG | |
CACAGCGATACAGGACAG | AGTGGGCCAAGATTGTACG | ||
基因Gene | 5'扩增引物 5' primers | 3'扩增引物3' primers | |
实时荧光定量PCR RT-qPCR | EfOctβ1R | CCTGTGGGTATTGTCGTTCGTC | CGTAGCCTGCGTCGTTGGTTA |
EfOctβ2R | TATCGTTCTGGGTGCCTGGAG | CGGACATAGATGCTGCGTAAC | |
EfEF1a | CCGCCAACATCACTACTGAAGT | GCAACATAACCACGACGCAAT |
蛋白Protein | 登录号Accession number | 物种Species | 蛋白Protein | 登录号Accession number | 物种Species |
---|---|---|---|---|---|
ClOctβ1R | XP_014241423.1 | Cimex lectularius | TdOctβ1R | XP_037947986.1 | Teleopsis dalmanni |
NlOctβ1R | ATY68966.1 | Nilaparvata lugens | DbOctβ1R | XP_017849616.1 | Drosophila busckii |
CsOctβ1R | XP_033609947.1 | Cryptotermes secundus | DhOctβ1R | XP_023175715.1 | Drosophila hydei |
ZnOctβ1R | XP_021920523.1 | Zootermopsis nevadensis | DwOctβ1R | EDW85043.1 | Drosophila willistoni |
ObOctβ1R | XP_011349656.1 | Ooceraea biroi | DmOctβ1R | XP_002000086.1 | Drosophila mojavensis |
CcOctβ1R | XP_015594309.1 | Cephus cinctus | DaOctβ1R | XP_017855803.1 | Drosophila arizonae |
TcOctβ1R | NP_001280514.1 | Tribolium castaneum | DvOctβ1R | XP_002054844.1 | Drosophila virilis |
PxOctβ1R | XP_013165427.1 | Papilio xuthus | DiOctβ1R | XP_034488357.1 | Drosophila innubila |
PmOctβ1R | XP_014360567.1 | Papilio machaon | DeOctβ1R | XP_001982264.1 | Drosophila erecta |
GmOctβ1R | XP_031764033.1 | Galleria mellonella | DsOctβ1R | XP_034665037.1 | Drosophila subobscura |
TnOctβ1R | XP_026729368.1 | Trichoplusia ni | DtOctβ1R | XP_017007379.1 | Drosophila takahashii |
VtOctβ1R | XP_026501717.1 | Vanessa tameamea | DgOctβ1R | XP_034127656.1 | Drosophila guanche |
DpOctβ1R | XP_032523541.1 | Danaus plexippus plexippus | DyOctβ1R | XP_002098302.1 | Drosophila yakuba |
NvOctβ1R | NP_001317478.1 | Nicrophorus vespilloides | CfOctβ1R | XP_026475745.1 | Ctenocephalides felis |
HaOctβ1R | XP_021187626.1 | Helicoverpa armigera | HhOctβ2R | XP_014285375.1 | Halyomorpha halys |
PrOctβ1R | XP_022123444.1 | Pieris rapae | ClOctβ2R | XP_014254146.1 | Cimex lectularius |
BaOctβ1R | XP_023952781.1 | Bicyclus anynana | RpOctβ2R | ATI14906.1 | Rhodnius prolixus |
BmOctβ1R | XP_004922133.2 | Bombyx mori | NlOctβ2R | ASA47149.1 | Nilaparvata lugens |
AtOctβ1R | XP_019864623.1 | Aethina tumida | ZnOctβ2R | XP_021931096.1 | Zootermopsis nevadensis |
MhOctβ1R | XP_034837322.1 | Maniola hyperantus | CsOctβ2R | XP_023721065.1 | Cryptotermes secundus |
HkOctβ1R | XP_026322929.1 | Hyposmocoma kahamanoa | LsOctβ2R | RZF43302.1 | Laodelphax striatellus |
LsOctβ1R | VVC99498.1 | Leptidea sinapis | BgOctβ2R | PSN51957.1 | Blattella germanica |
TpOctβ1R | XP_034238788.1 | Thrips palmi | TsOctβ2R | CAD7258263.1 | Timema shepardi |
FaOctβ1R | XP_011312392.1 | Fopius arisanus | TdOctβ2R | CAD7194428.1 | Timema douglasi |
ArOctβ1R | XP_020712470.1 | Athalia rosae | TmOctβ2R | CAD7423498.1 | Timema monikensis |
OfOctβ1R | XP_028167767.1 | Ostrinia furnacalis | TbOctβ2R | CAD7437944.1 | Timema bartmani |
PhOctβ1R | XP_002422997.1 | Pediculus humanus corporis | TpOctβ2R | XP_034233045.1 | Thrips palmi |
MsOctβ1R | XP_037296074.1 | Manduca sexta | NvOctβ2R | XP_017768337.1 | Nicrophorus vespilloides |
HsOctβ1R | XP_011135559.1 | Harpegnathos saltator | DqOctβ2R | XP_014477130.1 | Dinoponera quadriceps |
FoOctβ1R | XP_026276499.1 | Frankliniella occidentalis | CfOctβ2R | XP_011263103.1 | Camponotus floridanus |
PgOctβ1R | XP_020284449.1 | Pseudomyrmex gracilis | SiOctβ2R | XP_011166273.1 | Solenopsis invicta |
DqOctβ1R | XP_014467276.1 | Dinoponera quadriceps | LhOctβ2R | XP_012226543.1 | Linepithema humile |
SlOctβ1R | XP_022827519.1 | Spodoptera litura | CcOctβ2R | XP_018400970.1 | Cyphomyrmex costatus |
ApOctβ1R | XP_025836197.1 | Agrilus planipennis | ObOctβ2R | XP_032684807.1 | Odontomachus brunneus |
BtOctβ1R | XP_018906333.1 | Bemisia tabaci | TcOctβ2R | XP_024891919.1 | Temnothorax curvispinosus |
AcOctβ1R | KYM86550.1 | Atta colombica | TzOctβ2R | XP_018309105.1 | Trachymyrmex zeteki |
HiOctβ1R | XP_037913033.1 | Hermetia illucens | ApOctβ2R | XP_018331973.1 | Agrilus planipennis |
TsOctβ1R | KYN34595.1 | Trachymyrmex septentrionalis | WaOctβ2R | XP_011702425.1 | Wasmannia auropunctata |
DcOctβ1R | XP_008477582.2 | Diaphorina citri | MpOctβ2R | XP_012535778.1 | Monomorium pharaonis |
MpOctβ1R | XP_022179990.1 | Myzus persicae | FeOctβ2R | XP_029669314.1 | Formica exsecta |
DnOctβ1R | XP_015364764.1 | Diuraphis noxia | AfOctβ2R | XP_003694478.1 | Apis florea |
RmOctβ1R | XP_026809104.1 | Rhopalosiphum maidis | PbOctβ2R | XP_011643320.1 | Pogonomyrmex barbatus |
SfOctβ1R | XP_025409207.1 | Sipha flava | BtOctβ2R | XP_033219965.1 | Belonocnema treatae |
ScOctβ1R | XP_013099911.1 | Stomoxys calcitrans | NcOctβ2R | QID89710.1 | Nephotettix cincticeps |
GfOctβ1R | XP_037885413.1 | Glossina fuscipes | AdOctβ2R | XP_006621346.1 | Apis dorsata |
AcOctβ2R | XP_016908111.1 | Apis cerana | MgOctβ2R | XP_033333269.1 | Megalopta genalis |
DnOctβ2R | XP_015429562.1 | Dufourea novaeangliae | BvOctβ2R | XP_033362706.1 | Bombus vosnesenskii |
MrOctβ2R | XP_003701983.1 | Megachile rotundata | OaOctβ2R | XP_012281474.1 | Orussus abietinus |
DcOctβ2R | XP_026679423.1 | Diaphorina citri | CiOctβ2R | XP_034939572.1 | Chelonus insularis |
HlOctβ2R | XP_017790436.1 | Habropoda laboriosa | CsOctβ2R | XP_011504152.1 | Ceratosolen solmsi marchali |
AmOctβ2R | XP_006558130.1 | Apis mellifera | ArOctβ2R | XP_012261826.1 | Athalia rosae |
PcOctβ2R | XP_014616065.1 | Polistes canadensis | OtOctβ2R | XP_022918261.1 | Onthophagus taurus |
AeOctβ2R | XP_011065978.1 | Acromyrmex echinatior | LyOctβ2R | KAF5287830.1 | Lamprigera yunnana |
NmOctβ2R | XP_031833977.1 | Nomia melanderi | VmOctβ2R | XP_035736463.1 | Vespa mandarini |
EmOctβ2R | XP_017763346.1 | Eufriesea mexicana | FaOctβ2R | XP_011300779.1 | Fopius arisanus |
PdOctβ2R | XP_015180910.1 | Polistes dominula | MsOctβ2R | XP_025204042.1 | Melanaphis sacchari |
NfOctβ2R | XP_029169862.1 | Nylanderia fulva | RmOctβ2R | XP_026809287.1 | Rhopalosiphum maidis |
HsOctβ2R | XP_011140150.2 | Harpegnathos saltator | DvOctβ2R | XP_028130069.1 | Diabrotica virgifera virgifera |
ObOctβ2R | XP_029045046.1 | Osmia bicornis bicornis | LdOctβ2R | XP_023015134.1 | Leptinotarsa decemlineata |
VeOctβ2R | XP_011876313.1 | Vollenhovia emeryi | DaOctβ2R | XP_015109333.1 | Diachasma alloeum |
OlOctβ2R | XP_034187840.1 | Osmia lignaria | SfOctβ2R | XP_025423429.1 | Sipha flava |
BiOctβ2R | XP_003488300.1 | Bombus impatiens | AtOctβ2R | XP_019867648.1 | Aethina tumida |
MdOctβ2R | XP_008548223.1 | Microplitis demolitor |
Table 2 List sequence information used in phylogenetic analysis
蛋白Protein | 登录号Accession number | 物种Species | 蛋白Protein | 登录号Accession number | 物种Species |
---|---|---|---|---|---|
ClOctβ1R | XP_014241423.1 | Cimex lectularius | TdOctβ1R | XP_037947986.1 | Teleopsis dalmanni |
NlOctβ1R | ATY68966.1 | Nilaparvata lugens | DbOctβ1R | XP_017849616.1 | Drosophila busckii |
CsOctβ1R | XP_033609947.1 | Cryptotermes secundus | DhOctβ1R | XP_023175715.1 | Drosophila hydei |
ZnOctβ1R | XP_021920523.1 | Zootermopsis nevadensis | DwOctβ1R | EDW85043.1 | Drosophila willistoni |
ObOctβ1R | XP_011349656.1 | Ooceraea biroi | DmOctβ1R | XP_002000086.1 | Drosophila mojavensis |
CcOctβ1R | XP_015594309.1 | Cephus cinctus | DaOctβ1R | XP_017855803.1 | Drosophila arizonae |
TcOctβ1R | NP_001280514.1 | Tribolium castaneum | DvOctβ1R | XP_002054844.1 | Drosophila virilis |
PxOctβ1R | XP_013165427.1 | Papilio xuthus | DiOctβ1R | XP_034488357.1 | Drosophila innubila |
PmOctβ1R | XP_014360567.1 | Papilio machaon | DeOctβ1R | XP_001982264.1 | Drosophila erecta |
GmOctβ1R | XP_031764033.1 | Galleria mellonella | DsOctβ1R | XP_034665037.1 | Drosophila subobscura |
TnOctβ1R | XP_026729368.1 | Trichoplusia ni | DtOctβ1R | XP_017007379.1 | Drosophila takahashii |
VtOctβ1R | XP_026501717.1 | Vanessa tameamea | DgOctβ1R | XP_034127656.1 | Drosophila guanche |
DpOctβ1R | XP_032523541.1 | Danaus plexippus plexippus | DyOctβ1R | XP_002098302.1 | Drosophila yakuba |
NvOctβ1R | NP_001317478.1 | Nicrophorus vespilloides | CfOctβ1R | XP_026475745.1 | Ctenocephalides felis |
HaOctβ1R | XP_021187626.1 | Helicoverpa armigera | HhOctβ2R | XP_014285375.1 | Halyomorpha halys |
PrOctβ1R | XP_022123444.1 | Pieris rapae | ClOctβ2R | XP_014254146.1 | Cimex lectularius |
BaOctβ1R | XP_023952781.1 | Bicyclus anynana | RpOctβ2R | ATI14906.1 | Rhodnius prolixus |
BmOctβ1R | XP_004922133.2 | Bombyx mori | NlOctβ2R | ASA47149.1 | Nilaparvata lugens |
AtOctβ1R | XP_019864623.1 | Aethina tumida | ZnOctβ2R | XP_021931096.1 | Zootermopsis nevadensis |
MhOctβ1R | XP_034837322.1 | Maniola hyperantus | CsOctβ2R | XP_023721065.1 | Cryptotermes secundus |
HkOctβ1R | XP_026322929.1 | Hyposmocoma kahamanoa | LsOctβ2R | RZF43302.1 | Laodelphax striatellus |
LsOctβ1R | VVC99498.1 | Leptidea sinapis | BgOctβ2R | PSN51957.1 | Blattella germanica |
TpOctβ1R | XP_034238788.1 | Thrips palmi | TsOctβ2R | CAD7258263.1 | Timema shepardi |
FaOctβ1R | XP_011312392.1 | Fopius arisanus | TdOctβ2R | CAD7194428.1 | Timema douglasi |
ArOctβ1R | XP_020712470.1 | Athalia rosae | TmOctβ2R | CAD7423498.1 | Timema monikensis |
OfOctβ1R | XP_028167767.1 | Ostrinia furnacalis | TbOctβ2R | CAD7437944.1 | Timema bartmani |
PhOctβ1R | XP_002422997.1 | Pediculus humanus corporis | TpOctβ2R | XP_034233045.1 | Thrips palmi |
MsOctβ1R | XP_037296074.1 | Manduca sexta | NvOctβ2R | XP_017768337.1 | Nicrophorus vespilloides |
HsOctβ1R | XP_011135559.1 | Harpegnathos saltator | DqOctβ2R | XP_014477130.1 | Dinoponera quadriceps |
FoOctβ1R | XP_026276499.1 | Frankliniella occidentalis | CfOctβ2R | XP_011263103.1 | Camponotus floridanus |
PgOctβ1R | XP_020284449.1 | Pseudomyrmex gracilis | SiOctβ2R | XP_011166273.1 | Solenopsis invicta |
DqOctβ1R | XP_014467276.1 | Dinoponera quadriceps | LhOctβ2R | XP_012226543.1 | Linepithema humile |
SlOctβ1R | XP_022827519.1 | Spodoptera litura | CcOctβ2R | XP_018400970.1 | Cyphomyrmex costatus |
ApOctβ1R | XP_025836197.1 | Agrilus planipennis | ObOctβ2R | XP_032684807.1 | Odontomachus brunneus |
BtOctβ1R | XP_018906333.1 | Bemisia tabaci | TcOctβ2R | XP_024891919.1 | Temnothorax curvispinosus |
AcOctβ1R | KYM86550.1 | Atta colombica | TzOctβ2R | XP_018309105.1 | Trachymyrmex zeteki |
HiOctβ1R | XP_037913033.1 | Hermetia illucens | ApOctβ2R | XP_018331973.1 | Agrilus planipennis |
TsOctβ1R | KYN34595.1 | Trachymyrmex septentrionalis | WaOctβ2R | XP_011702425.1 | Wasmannia auropunctata |
DcOctβ1R | XP_008477582.2 | Diaphorina citri | MpOctβ2R | XP_012535778.1 | Monomorium pharaonis |
MpOctβ1R | XP_022179990.1 | Myzus persicae | FeOctβ2R | XP_029669314.1 | Formica exsecta |
DnOctβ1R | XP_015364764.1 | Diuraphis noxia | AfOctβ2R | XP_003694478.1 | Apis florea |
RmOctβ1R | XP_026809104.1 | Rhopalosiphum maidis | PbOctβ2R | XP_011643320.1 | Pogonomyrmex barbatus |
SfOctβ1R | XP_025409207.1 | Sipha flava | BtOctβ2R | XP_033219965.1 | Belonocnema treatae |
ScOctβ1R | XP_013099911.1 | Stomoxys calcitrans | NcOctβ2R | QID89710.1 | Nephotettix cincticeps |
GfOctβ1R | XP_037885413.1 | Glossina fuscipes | AdOctβ2R | XP_006621346.1 | Apis dorsata |
AcOctβ2R | XP_016908111.1 | Apis cerana | MgOctβ2R | XP_033333269.1 | Megalopta genalis |
DnOctβ2R | XP_015429562.1 | Dufourea novaeangliae | BvOctβ2R | XP_033362706.1 | Bombus vosnesenskii |
MrOctβ2R | XP_003701983.1 | Megachile rotundata | OaOctβ2R | XP_012281474.1 | Orussus abietinus |
DcOctβ2R | XP_026679423.1 | Diaphorina citri | CiOctβ2R | XP_034939572.1 | Chelonus insularis |
HlOctβ2R | XP_017790436.1 | Habropoda laboriosa | CsOctβ2R | XP_011504152.1 | Ceratosolen solmsi marchali |
AmOctβ2R | XP_006558130.1 | Apis mellifera | ArOctβ2R | XP_012261826.1 | Athalia rosae |
PcOctβ2R | XP_014616065.1 | Polistes canadensis | OtOctβ2R | XP_022918261.1 | Onthophagus taurus |
AeOctβ2R | XP_011065978.1 | Acromyrmex echinatior | LyOctβ2R | KAF5287830.1 | Lamprigera yunnana |
NmOctβ2R | XP_031833977.1 | Nomia melanderi | VmOctβ2R | XP_035736463.1 | Vespa mandarini |
EmOctβ2R | XP_017763346.1 | Eufriesea mexicana | FaOctβ2R | XP_011300779.1 | Fopius arisanus |
PdOctβ2R | XP_015180910.1 | Polistes dominula | MsOctβ2R | XP_025204042.1 | Melanaphis sacchari |
NfOctβ2R | XP_029169862.1 | Nylanderia fulva | RmOctβ2R | XP_026809287.1 | Rhopalosiphum maidis |
HsOctβ2R | XP_011140150.2 | Harpegnathos saltator | DvOctβ2R | XP_028130069.1 | Diabrotica virgifera virgifera |
ObOctβ2R | XP_029045046.1 | Osmia bicornis bicornis | LdOctβ2R | XP_023015134.1 | Leptinotarsa decemlineata |
VeOctβ2R | XP_011876313.1 | Vollenhovia emeryi | DaOctβ2R | XP_015109333.1 | Diachasma alloeum |
OlOctβ2R | XP_034187840.1 | Osmia lignaria | SfOctβ2R | XP_025423429.1 | Sipha flava |
BiOctβ2R | XP_003488300.1 | Bombus impatiens | AtOctβ2R | XP_019867648.1 | Aethina tumida |
MdOctβ2R | XP_008548223.1 | Microplitis demolitor |
基因Gene | 开放阅读框ORF/bp | 蛋白Protein/aa | 蛋白分子量MW/kD | 等电点 pI | 亲水系数GRAVY |
---|---|---|---|---|---|
EfOctβ1R | 1 245 | 414 | 47.34 | 9.02 | 0.194 |
EfOctβ2R | 1 230 | 409 | 46.67 | 8.76 | 0.260 |
Table 3 List of EfOctβ1R and EfOctβ2R in Eocanthecona furcellata
基因Gene | 开放阅读框ORF/bp | 蛋白Protein/aa | 蛋白分子量MW/kD | 等电点 pI | 亲水系数GRAVY |
---|---|---|---|---|---|
EfOctβ1R | 1 245 | 414 | 47.34 | 9.02 | 0.194 |
EfOctβ2R | 1 230 | 409 | 46.67 | 8.76 | 0.260 |
Fig. 1 Amino acid sequence alignment of EfOctβ1R in E. furrellata and orthologous receptors from Nilaparvata lugens(NlOctβ1R,ATY68966.1),Tribolium castaneum(TcOctβ1R,NP_001280514.1)and Drosophila melanogaster(DmOctβ1R,NP_001163690.1) The seven transmembrane regions are predicted by TM1-TM7. Potential N-glycosylation sites and protein kinases binding sites by PKC are labelled by○and●,respectively. The filled arrows mark as cysteine residue ligand binding sites are marked with★. The FxxxWxP motif in TM6 is indicated by◇,The same below
Fig. 2 Amino acid sequence alignment of EfOctβ2R and orthologous receptors from Cimex lectularius(ClOctβ2R,XP_014254146.1),Nilaparvata lugens(NlOctβ2R,ASA47149.1)and Tribolium castaneum(TcOctβ2R,NP_001280501.1)
Fig. 4 Expression pattern of EfOctβ1R and EfOctβ2 R in E. furcellata Data are represented as the mean±SD from four independent experiments with EfEF1a as the housekeeping gene,the same below
Fig. 5 Change in the gene expressions of EfOctβ1R and EfOctβ2R in E. furellata treated with sublethal doses of efficient r-cyhalothrin and chlorpyrifos
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