The objective of this study is to express the carbohydrate recognition domain （CRD） of the asialoglycoprotein receptor （ASGPR） H1 and H2 subunits of Marmota himalayan in vitro, and develop polyclonal antibodies ag...The objective of this study is to express the carbohydrate recognition domain （CRD） of the asialoglycoprotein receptor （ASGPR） H1 and H2 subunits of Marmota himalayan in vitro, and develop polyclonal antibodies against the recombinant proteins. RT-PCR was used to amplify ASGPR CRDH1 and CRDH2 from the liver tissue of Marmota himalayan. The products of amplification were subcloned into prokaryotic expression vector pRSET-B, and expressed in E.coli BL21（DE3）plysS. The recombinant proteins were purified using Ni-NTA spin column. The purified proteins were inoculated into BALB/c mice to develop polyclonal antibodies. The sensitivity and specificity of antibodies were evaluated by enzyme-linked immunosorbent assay （ELISA）, Western blotting and immunohistochemical staining （IHC）. The polyclonal antibodies showed high sensitivity and specificity against both denaturated and native ASGPR proteins. We successfully amplified and expressed the ASGPR CRDs of Marmota himalayan. The nucleic sequences of ASGPR CRDH1 and CRDH2 of Marmota himalayan have been submitted to Genbank and the sequence ID are DQ 845465 and DQ845466, respectively. The proteins and antibodies prepared can be used for targeting gene therapy in a new animal model-Marrnota himalayan—— for the research of infectious diseases of hepatitis viruses and liver cancer treatment.展开更多
Few studies have examined the succession of plant communities in the alpine zone.Studying the succession of plant communities is helpful to understand how species diversity is formed and maintained.In this study,we us...Few studies have examined the succession of plant communities in the alpine zone.Studying the succession of plant communities is helpful to understand how species diversity is formed and maintained.In this study,we used species inventories,a molecular phylogeny,and trait data to detect patterns of phylogenetic and functional community structure in successional plant communities growing on the mounds of Himalayan marmots(Marmota himalayana)on the southeast edge of the Qinghai-Tibet Plateau.We found that phylogenetic and functional diversities of plant communities on marmot mounds tended to cluster during the early to medium stages of succession,then trended toward overdispersion from medium to late stages.Alpine species in early and late stages of succession were phylogenetically and functionally overdispersed,suggesting that such communities were assembled mainly through species interactions,especially competition.At the medium and late stages of succession,alpine communities growing on marmot mounds were phylogenetically and functionally clustered,implying that the communities were primarily structured by environmental filtering.During the medium and late stages of succession the phylogenetic and functional structures of plant communities on marmot mounds differed significantly from those on neighboring sites.Our results indicate that environmental filtering and species interactions can change plant community composition at different successional stages.Assembly of plant communities on marmot mounds was promoted by a combination of traits that may provide advantages for survival and adaptation during periods of environmental change.展开更多
基金This project was supported by grants from National Natural Sciences Foundation of China (No 30571646)the Na-tional Basic Research Program (Program 973, No 2005CB522901)
文摘The objective of this study is to express the carbohydrate recognition domain （CRD） of the asialoglycoprotein receptor （ASGPR） H1 and H2 subunits of Marmota himalayan in vitro, and develop polyclonal antibodies against the recombinant proteins. RT-PCR was used to amplify ASGPR CRDH1 and CRDH2 from the liver tissue of Marmota himalayan. The products of amplification were subcloned into prokaryotic expression vector pRSET-B, and expressed in E.coli BL21（DE3）plysS. The recombinant proteins were purified using Ni-NTA spin column. The purified proteins were inoculated into BALB/c mice to develop polyclonal antibodies. The sensitivity and specificity of antibodies were evaluated by enzyme-linked immunosorbent assay （ELISA）, Western blotting and immunohistochemical staining （IHC）. The polyclonal antibodies showed high sensitivity and specificity against both denaturated and native ASGPR proteins. We successfully amplified and expressed the ASGPR CRDs of Marmota himalayan. The nucleic sequences of ASGPR CRDH1 and CRDH2 of Marmota himalayan have been submitted to Genbank and the sequence ID are DQ 845465 and DQ845466, respectively. The proteins and antibodies prepared can be used for targeting gene therapy in a new animal model-Marrnota himalayan—— for the research of infectious diseases of hepatitis viruses and liver cancer treatment.
基金the National Natural Science Foundation of China(NSFC)(Grant No.31560063)Applied Basic Research Program of Yunnan Province,China(2018FB067).
文摘Few studies have examined the succession of plant communities in the alpine zone.Studying the succession of plant communities is helpful to understand how species diversity is formed and maintained.In this study,we used species inventories,a molecular phylogeny,and trait data to detect patterns of phylogenetic and functional community structure in successional plant communities growing on the mounds of Himalayan marmots(Marmota himalayana)on the southeast edge of the Qinghai-Tibet Plateau.We found that phylogenetic and functional diversities of plant communities on marmot mounds tended to cluster during the early to medium stages of succession,then trended toward overdispersion from medium to late stages.Alpine species in early and late stages of succession were phylogenetically and functionally overdispersed,suggesting that such communities were assembled mainly through species interactions,especially competition.At the medium and late stages of succession,alpine communities growing on marmot mounds were phylogenetically and functionally clustered,implying that the communities were primarily structured by environmental filtering.During the medium and late stages of succession the phylogenetic and functional structures of plant communities on marmot mounds differed significantly from those on neighboring sites.Our results indicate that environmental filtering and species interactions can change plant community composition at different successional stages.Assembly of plant communities on marmot mounds was promoted by a combination of traits that may provide advantages for survival and adaptation during periods of environmental change.
