Effects of different enucleation methods on developmental potency of pig handmade clone（HMC）reconstructed embryo was investigated in the paper.We compared enucleation efficiency of blind cut method,first polar body...Effects of different enucleation methods on developmental potency of pig handmade clone（HMC）reconstructed embryo was investigated in the paper.We compared enucleation efficiency of blind cut method,first polar body（Pb1）positioning method and demecolcine（DM）assisted enucleation,as well as their effects on development of HMC reconstructed embryos.The results showed that overall enucleation efficiency of Pb1 positioning method was significantly higher than that of blind cut method（P〈0.05）.The protuberance rate and overall enucleation efficiency of 0.4μg/mL DM treated group for 60 min was significantly higher than that of other concentrations and time treatment groups（P〈0.05）.For effects on development of HMC reconstructed embryos,there was no significant difference between DM-assisted enucleation and Pb1 positioning method.In conclusion,appropriate addition of DM could enhance enucleation efficiency of HMC,which had no significant influence on developmental potency of reconstructed embryos.展开更多
Flooding is the most famous technique for locating contents in unstructured P2P networks. Recently traditional flooding has been replaced by more efficient dynamic query （DQ） and different variants of such algorithm...Flooding is the most famous technique for locating contents in unstructured P2P networks. Recently traditional flooding has been replaced by more efficient dynamic query （DQ） and different variants of such algorithms. Dynamic query is a new flooding technique which could estimate a proper time-to-live （TTL） value for a query flooding by estimating the popularity of the searched files, and retrieve sufficient results under controlled flooding range for reducing network traffic. However, all DQ-like search algorithms are ＂blind＂ so that a large amount of redundant messages are caused. In this paper, we proposed a new search scheme, called Immune Search Scheme （ISS）, to cope with this problem. In ISS, an immune systems inspired concept of similarity-governed clone proliferation and mutation for query message movement is applied. Some assistant strategies, that is, shortcuts creation and peer traveling are incorporated into ISS to develop ＂immune memory＂ for improving search performance, which can make ISS not be blind but heuristic.展开更多
The yeast two-hybrid (Y2H) mating assay is a powerful method for detecting protein-protein interactions. Firstly, the gene of interest is cloned into specific Y2H vectors. Although multiple innovations in cloning meth...The yeast two-hybrid (Y2H) mating assay is a powerful method for detecting protein-protein interactions. Firstly, the gene of interest is cloned into specific Y2H vectors. Although multiple innovations in cloning methods were made in the past two decades, the conventional cloning method of restriction-enzyme (RE) digestion followed by ligation is still widely used. Unfortunately, many researchers, especially new-comers, often encounter difficulties in cloning a gene into a desired vector. Secondly, interaction between two proteins is commonly detected by growth of the diploids in specific media. This step takes about two weeks. Here, we describe improved cloning and detection procedures for the Y2H assay that accelerate the research progress. The changes in procedures involve running an agarose gel after the doubly digested vector and insert are ligated in the cloning step to determine the efficiency of RE digestion and ligation, and performing an additional replica-plating on plates for earlier assessment of interaction in the detection step. We show an example of Y2H interaction between Trs23 and Trs120 (respective subunits of TRAPP I and TRAPP II), as a proof of concept. By following the improved methods described here, the chances of successful cloning increased and the time for the whole Y2H experimental process is significantly shorter.展开更多
基金Supported by National Natural Science Foundation of China(31360544)
文摘Effects of different enucleation methods on developmental potency of pig handmade clone（HMC）reconstructed embryo was investigated in the paper.We compared enucleation efficiency of blind cut method,first polar body（Pb1）positioning method and demecolcine（DM）assisted enucleation,as well as their effects on development of HMC reconstructed embryos.The results showed that overall enucleation efficiency of Pb1 positioning method was significantly higher than that of blind cut method（P〈0.05）.The protuberance rate and overall enucleation efficiency of 0.4μg/mL DM treated group for 60 min was significantly higher than that of other concentrations and time treatment groups（P〈0.05）.For effects on development of HMC reconstructed embryos,there was no significant difference between DM-assisted enucleation and Pb1 positioning method.In conclusion,appropriate addition of DM could enhance enucleation efficiency of HMC,which had no significant influence on developmental potency of reconstructed embryos.
基金Supported by the National Natural Science Foundation of China (90604012)
文摘Flooding is the most famous technique for locating contents in unstructured P2P networks. Recently traditional flooding has been replaced by more efficient dynamic query （DQ） and different variants of such algorithms. Dynamic query is a new flooding technique which could estimate a proper time-to-live （TTL） value for a query flooding by estimating the popularity of the searched files, and retrieve sufficient results under controlled flooding range for reducing network traffic. However, all DQ-like search algorithms are ＂blind＂ so that a large amount of redundant messages are caused. In this paper, we proposed a new search scheme, called Immune Search Scheme （ISS）, to cope with this problem. In ISS, an immune systems inspired concept of similarity-governed clone proliferation and mutation for query message movement is applied. Some assistant strategies, that is, shortcuts creation and peer traveling are incorporated into ISS to develop ＂immune memory＂ for improving search performance, which can make ISS not be blind but heuristic.
文摘The yeast two-hybrid (Y2H) mating assay is a powerful method for detecting protein-protein interactions. Firstly, the gene of interest is cloned into specific Y2H vectors. Although multiple innovations in cloning methods were made in the past two decades, the conventional cloning method of restriction-enzyme (RE) digestion followed by ligation is still widely used. Unfortunately, many researchers, especially new-comers, often encounter difficulties in cloning a gene into a desired vector. Secondly, interaction between two proteins is commonly detected by growth of the diploids in specific media. This step takes about two weeks. Here, we describe improved cloning and detection procedures for the Y2H assay that accelerate the research progress. The changes in procedures involve running an agarose gel after the doubly digested vector and insert are ligated in the cloning step to determine the efficiency of RE digestion and ligation, and performing an additional replica-plating on plates for earlier assessment of interaction in the detection step. We show an example of Y2H interaction between Trs23 and Trs120 (respective subunits of TRAPP I and TRAPP II), as a proof of concept. By following the improved methods described here, the chances of successful cloning increased and the time for the whole Y2H experimental process is significantly shorter.
