生物学杂志
生物學雜誌
생물학잡지
JOURNAL OF BIOLOGY
2015年
1期
1-5
,共5页
徐泽文%杨筱珍%黄坚%李彤%杨志刚%王春%成永旭
徐澤文%楊篠珍%黃堅%李彤%楊誌剛%王春%成永旭
서택문%양소진%황견%리동%양지강%왕춘%성영욱
CODEHOP%中华绒螯蟹%5-ht1r%5-ht2r%克隆
CODEHOP%中華絨螯蟹%5-ht1r%5-ht2r%剋隆
CODEHOP%중화융오해%5-ht1r%5-ht2r%극륭
CODEHOP%Eir ocheir is nensis%5h-t1%r5-ht2r%cloning
根据已报道的中华绒螯蟹近缘物种的5-ht1 r和5-ht2 r氨基酸序列保守区,利用CODEHOP设计了两对简并引物,采用RT-PCR法扩增得到了中华绒螯蟹5-ht1 r和5-ht2 r部分片段并克隆到pMD-19T载体。测序结果表明扩增到的中华绒螯蟹5-ht1 r和5-ht2 r片段长分别为366 bp和177 bp,编码121和58个氨基酸。经BLASTp分析显示,中华绒螯蟹5-ht1r与罗氏沼虾(Macrobrachium rosenbergii)和克氏原螯虾(Procambarus clarkii)的5-ht1r同源性高达98%,而中华绒螯蟹5-HT2R与断沟龙虾(Panulirus interruptus)和克氏原螯虾(Procambarus clarkii)5-ht2r同源性高达95%。
根據已報道的中華絨螯蟹近緣物種的5-ht1 r和5-ht2 r氨基痠序列保守區,利用CODEHOP設計瞭兩對簡併引物,採用RT-PCR法擴增得到瞭中華絨螯蟹5-ht1 r和5-ht2 r部分片段併剋隆到pMD-19T載體。測序結果錶明擴增到的中華絨螯蟹5-ht1 r和5-ht2 r片段長分彆為366 bp和177 bp,編碼121和58箇氨基痠。經BLASTp分析顯示,中華絨螯蟹5-ht1r與囉氏沼蝦(Macrobrachium rosenbergii)和剋氏原螯蝦(Procambarus clarkii)的5-ht1r同源性高達98%,而中華絨螯蟹5-HT2R與斷溝龍蝦(Panulirus interruptus)和剋氏原螯蝦(Procambarus clarkii)5-ht2r同源性高達95%。
근거이보도적중화융오해근연물충적5-ht1 r화5-ht2 r안기산서렬보수구,이용CODEHOP설계료량대간병인물,채용RT-PCR법확증득도료중화융오해5-ht1 r화5-ht2 r부분편단병극륭도pMD-19T재체。측서결과표명확증도적중화융오해5-ht1 r화5-ht2 r편단장분별위366 bp화177 bp,편마121화58개안기산。경BLASTp분석현시,중화융오해5-ht1r여라씨소하(Macrobrachium rosenbergii)화극씨원오하(Procambarus clarkii)적5-ht1r동원성고체98%,이중화융오해5-HT2R여단구룡하(Panulirus interruptus)화극씨원오하(Procambarus clarkii)5-ht2r동원성고체95%。
According to the conserved amino acid sequences of known 5-Hydroxytryptamine receptors , CODEHOP ( Con-sensus Degenerate Hybrid Oligonucleotide Primers ) software was used to design degenerate primers to obtain partial cDNA fragments of 5-HT1 and 5-HT2 receptors (5-ht1r and 5-ht2r) in Chinese mitten crab (Eriocheir sinensis).The fragments of 5-ht1 r and 5-ht2 r were amplified by reverse transcription polymerase chain reaction ( RT-PCR) and then cloned into pMD-19T vector.The sequencing results revealed that 5-ht1 r and 5-ht2 r fragments contain 366 and 177 base pairs, which en-code 121 and 58 amino acids respectively .BLAST analysis indicated that the amino acid homology of 5-ht1 r was 98%sim-ilar to Macrobrachium rosenbergii and Procambarus clarkia, and the 5-ht2 r was most similar to Panulirus interruptus and Procambarus clarkia (95%) .