采用SMART(switching metchanism at 5’end of RNA transcript)技术构建缢蛏cDNA基因文库。并对cDNA文库的滴度,重组率和插入片段的大小进行了检测。结果表明,该cDNA文库的滴度为5.50×104cfu/ml,重组率为92.5%,插入片段的长度大多...采用SMART(switching metchanism at 5’end of RNA transcript)技术构建缢蛏cDNA基因文库。并对cDNA文库的滴度,重组率和插入片段的大小进行了检测。结果表明,该cDNA文库的滴度为5.50×104cfu/ml,重组率为92.5%,插入片段的长度大多在1kb以上。对478个克隆子进行了测序,共得到430个有效序列,其中166个序列无同源性,264条序列具有同源性。从中得到肌动蛋白(actin)cDNA的全长1538bp,开放阅读框(ORF)为1131bp,编码376个氨基酸,其预测蛋白的分子量为41.78kD,等电点为5.30。该氨基酸序列与其它物种的同源性大都在90%以上。展开更多
To construct an effective circular mix-culture model and investigate its culture capacity,we established one small and one large systems,with white shrimp(Litopenaeus vannamei)and razor clam(Sinonovacula constricta)cu...To construct an effective circular mix-culture model and investigate its culture capacity,we established one small and one large systems,with white shrimp(Litopenaeus vannamei)and razor clam(Sinonovacula constricta)cultured in separate ponds.The culture water from the L.vannamei was pumped to the S.constricta,and the culture water from the S.constricta overflowed back into the L.vannamei via gravity.In trial I,we tested four culture densities(groups 1–4),and monitored water quality,growth indices,and digestive and immune enzyme activities.From the results,the nitrogen and phosphorus levels generally increased then declined after 56 days,and were lower in groups 1 and 2.The specific growth rate of group 2 was the highest.After 56 days,activities of four digestive enzymes were increased in group 2,and lysozyme activity was significantly decreased in S.constricta in groups 1–4;Alkaline phosphatase activity of L.vannamei was increased in group 3,but decreased in S.constricta in groups 2–4;Acid phosphatase activity was significantly higher in groups 1–3(P<0.05),while SOD and CAT activities were significantly elevated in group 2(P<0.05).Thus,we applied group 2 density for trial II.In trial II,nitrogen and phosphorus concentrations declined significantly during the latter stages,and growth indices were higher than the control(P<0.05).The yields of L.vannamei and S.constricta were significantly higher than the control(P<0.05).The results revealed a good circular mix-culture system with the optimal culture capacity for L.vannamei(40 ind m^(−2))and S.constricta(300 ind m^(−2)),which provided a reference for the future culture of them.展开更多
Peptidoglycan recognition protein(PGRP)plays a vital role in invertebrate innate immunity system as a specific pattern recognition receptor for peptidoglycan.Bivalves possess various PGRP systems for self-defense;howe...Peptidoglycan recognition protein(PGRP)plays a vital role in invertebrate innate immunity system as a specific pattern recognition receptor for peptidoglycan.Bivalves possess various PGRP systems for self-defense;however,it has not been characterized in razor clam Sinonovacula constricta.In this study,eight PGRP coding sequences were identified and analyzed from S.constricta genome,which are designated as ScPGRP-S1,ScPGRP-S2,ScPGRP-S3,ScPGRP-S4,ScPGRP-S5,ScPGRP-S6,ScPGRP-S7,ScPGRP-S8.The results of molecular evolutionary analyses showed that all eight ScPGRP genes were highly conserved and exhi-bited a typical PGRP/amidase_2 domain as PGRP genes in other mollusks.Moreover,the presence of signal peptides was predicted in ScPGRP-S2,ScPGRP-S3 and ScPRP-S6,while a transmembrane structure only existed in ScPGRP-S6.Notably,a tertiary struc-ture analysis indicated that no disulfide bond was observed in ScPGRP-S5 and ScPGRP-S7.The mRNA transcripts analysis of ScPGRPs revealed that the high expression patterns of ScPGRP-S1 and ScPGRP-S4 were found in mantle,adductor muscle and foot,while those of ScPGRP-S2,ScPGRP-S3 and ScPGRP-S6 were observed in hepatopancreas.Furthermore,the temporal expression profiles of ScPGRPs in the hepatopancreas were analyzed by qPCR<https://www.sciencedirect.com/topics/immunology-and-microbiology/real-time-polymerase-chain-reaction>after Gram-negative Vibrio parahaemolyticus and Gram-positive Staphylococcus aureus challenges.The mRNA expressions of ScPGRP-S2,ScPGRP-S3 and ScPGRP-S6 could be induced by V.pa-rahaemolyticus and S.aureus.Overall,our findings indicated that ScPGRPs were involved in the immune defense against invaders,which constituted a comprehensive understanding of the potential role of PGRP genes in S.constricta.展开更多
文摘采用SMART(switching metchanism at 5’end of RNA transcript)技术构建缢蛏cDNA基因文库。并对cDNA文库的滴度,重组率和插入片段的大小进行了检测。结果表明,该cDNA文库的滴度为5.50×104cfu/ml,重组率为92.5%,插入片段的长度大多在1kb以上。对478个克隆子进行了测序,共得到430个有效序列,其中166个序列无同源性,264条序列具有同源性。从中得到肌动蛋白(actin)cDNA的全长1538bp,开放阅读框(ORF)为1131bp,编码376个氨基酸,其预测蛋白的分子量为41.78kD,等电点为5.30。该氨基酸序列与其它物种的同源性大都在90%以上。
基金supported by the National Key Research and Development Plan(No.2019YFD0900400)the Ear marked Fund for Modern Agro-Industry Technology Research System,China(No.CARS-47)sponsored by the K.C.Wong Magna Fund in Ningbo University.
文摘To construct an effective circular mix-culture model and investigate its culture capacity,we established one small and one large systems,with white shrimp(Litopenaeus vannamei)and razor clam(Sinonovacula constricta)cultured in separate ponds.The culture water from the L.vannamei was pumped to the S.constricta,and the culture water from the S.constricta overflowed back into the L.vannamei via gravity.In trial I,we tested four culture densities(groups 1–4),and monitored water quality,growth indices,and digestive and immune enzyme activities.From the results,the nitrogen and phosphorus levels generally increased then declined after 56 days,and were lower in groups 1 and 2.The specific growth rate of group 2 was the highest.After 56 days,activities of four digestive enzymes were increased in group 2,and lysozyme activity was significantly decreased in S.constricta in groups 1–4;Alkaline phosphatase activity of L.vannamei was increased in group 3,but decreased in S.constricta in groups 2–4;Acid phosphatase activity was significantly higher in groups 1–3(P<0.05),while SOD and CAT activities were significantly elevated in group 2(P<0.05).Thus,we applied group 2 density for trial II.In trial II,nitrogen and phosphorus concentrations declined significantly during the latter stages,and growth indices were higher than the control(P<0.05).The yields of L.vannamei and S.constricta were significantly higher than the control(P<0.05).The results revealed a good circular mix-culture system with the optimal culture capacity for L.vannamei(40 ind m^(−2))and S.constricta(300 ind m^(−2)),which provided a reference for the future culture of them.
基金This work was supported by the National Key Research and Development Program of China(No.2018YFD0901405)the Zhejiang Major Program of Science and Technology(No.2016C02055-9)+1 种基金the Ningbo Major Project of Science and Technology(No.2019B10005)the China Agriculture Research System of MOF and MARA,National Marine Genetic Resource Center Program.
文摘Peptidoglycan recognition protein(PGRP)plays a vital role in invertebrate innate immunity system as a specific pattern recognition receptor for peptidoglycan.Bivalves possess various PGRP systems for self-defense;however,it has not been characterized in razor clam Sinonovacula constricta.In this study,eight PGRP coding sequences were identified and analyzed from S.constricta genome,which are designated as ScPGRP-S1,ScPGRP-S2,ScPGRP-S3,ScPGRP-S4,ScPGRP-S5,ScPGRP-S6,ScPGRP-S7,ScPGRP-S8.The results of molecular evolutionary analyses showed that all eight ScPGRP genes were highly conserved and exhi-bited a typical PGRP/amidase_2 domain as PGRP genes in other mollusks.Moreover,the presence of signal peptides was predicted in ScPGRP-S2,ScPGRP-S3 and ScPRP-S6,while a transmembrane structure only existed in ScPGRP-S6.Notably,a tertiary struc-ture analysis indicated that no disulfide bond was observed in ScPGRP-S5 and ScPGRP-S7.The mRNA transcripts analysis of ScPGRPs revealed that the high expression patterns of ScPGRP-S1 and ScPGRP-S4 were found in mantle,adductor muscle and foot,while those of ScPGRP-S2,ScPGRP-S3 and ScPGRP-S6 were observed in hepatopancreas.Furthermore,the temporal expression profiles of ScPGRPs in the hepatopancreas were analyzed by qPCR<https://www.sciencedirect.com/topics/immunology-and-microbiology/real-time-polymerase-chain-reaction>after Gram-negative Vibrio parahaemolyticus and Gram-positive Staphylococcus aureus challenges.The mRNA expressions of ScPGRP-S2,ScPGRP-S3 and ScPGRP-S6 could be induced by V.pa-rahaemolyticus and S.aureus.Overall,our findings indicated that ScPGRPs were involved in the immune defense against invaders,which constituted a comprehensive understanding of the potential role of PGRP genes in S.constricta.
