Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plasti...Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.展开更多
Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusin...Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retention signal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptide into ER, while ER retention signal can make the protein retained in the ER and its derivative protein body. ELISA test indicated that the accumulation level of foreign CpTI protein in sck transgenic tobacco (Nicotiana tabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four times higher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to the increased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be used to increase the accumulation level of foreign protein in transgenic plants and can be widely applied to other related research field in plant genetic engineering.展开更多
The vectors carrying the genes coding for the proteins of interest are of unpredictable efficiency in transgenic animals. The expression vector of mammary gland (pINGG) containing GCSF genomic DNA was injected into m...The vectors carrying the genes coding for the proteins of interest are of unpredictable efficiency in transgenic animals. The expression vector of mammary gland (pINGG) containing GCSF genomic DNA was injected into mouse mammary gland, and expression was detected in the milk of mice. The result showed that mammary gland injection method could provide a convenient transient system to confirm vector validity.展开更多
With the development of plant genetic engineering techniques, numerous genetically modified plants have been generated. At the same time, the technologies for detecting transgenic organisms get improved constantly, wh...With the development of plant genetic engineering techniques, numerous genetically modified plants have been generated. At the same time, the technologies for detecting transgenic organisms get improved constantly, which also promotes the scientific identification, evaluation and commercial cultivation of transgenic plants. In this review, we evaluate various detection methods for transgenic plants at the level of protein expression.展开更多
The promoter is a cis-acting element in regulating gene expression. A promoterless plasmid containing UidA gene was transformed into tritordeum by barmbadment. Histochemical analysis of various tissues in transgenic t...The promoter is a cis-acting element in regulating gene expression. A promoterless plasmid containing UidA gene was transformed into tritordeum by barmbadment. Histochemical analysis of various tissues in transgenic tritordeum was carried to examine tissue-specific expression of GUS(beta-glucuronidase) activity. The pollen-specific promoter was trapped and identified successfully in a transformant line. PCR(polymerase chain reaction) method was used to isolate this pollen-specific promoter. By sequencing and analyzing the amplified fragment from PCR, a part of UidA gene and a flanking sequence were obtained. Some essential elements of plant promoters were found in the sequence. To determine the function of it, the cloned fragment was fused with UidA gene, then cloned and transformed into Triticum durum. The transgenic plant transformed by this vector showed GUS expression only in pollen. Therefore a pollen-specific promoter was isolated successfully.展开更多
Cells are crowded microenvironments filled with macromolecules undergoing constant phys- ical and chemical interactions. The physicochemical makeup of the cells aff)cts various cellular responses, determines cell-cel...Cells are crowded microenvironments filled with macromolecules undergoing constant phys- ical and chemical interactions. The physicochemical makeup of the cells aff)cts various cellular responses, determines cell-cell interactions and influences cell decisions. Chemical and physical properties diff)r between cells and within cells. Moreover, these properties are subject to dynamic changes in response to environmental signals, which often demand adjustments in the chemical or physical states of intracellular molecules. Indeed, cellular responses such as gene expression rely on the faithful relay of information from the outside to the inside of the cell, a process terrned signal transduction. The signal often traverses a complex path across subcellular spaces with variable physical chemistry, sometimes even influencing it. Understanding the molecular states of such signaling molecules and their intracellular environments is vital to our understanding of the cell. Exploring such intricate spaces is possible today largely because of experimental and theoretical tools. Here, we focus on one tool that is commonly used in chemical physics studies light. We summarize recent work which uses light to both visualize the cellular environment and also control intracel- lular processes along the axis of signal transduction. We highlight recent accomplishments in optical microscopy and optogenetics, an emerging experimental strategy which utilizes light to control the molecular processes in live cells. We believe that optogenetics lends un- precedented spatiotemporal precision to the manipulation of physicochemical properties in biological contexts. We hope to use this work to demonstrate new opportunities for chemical physicists who are interested in pursuing biological and biomedical questions.展开更多
Recent deep sequencing surveys of mammalian genomes have unexpectedly revealed pervasive and complex transcription and identified tens of thousands of RNA transcripts that do not code for proteins. These non-coding RN...Recent deep sequencing surveys of mammalian genomes have unexpectedly revealed pervasive and complex transcription and identified tens of thousands of RNA transcripts that do not code for proteins. These non-coding RNAs(nc RNAs) highlight the central role of RNA in gene regulation. nc RNAs are arbitrarily divided into two main groups: The first includes small RNAs, such as mi RNAs, pi RNAs, and endogenous si RNAs, that usually range from 20 to 30 nt, while the second group includes long non-coding RNAs(lnc RNAs), which are typically more than 200 nt in length. These nc RNAs were initially thought to merely regulate gene expression at the post-transcriptional level, but recent studies have indicated that nc RNAs, especially lnc RNAs, are extensively associated with diverse chromatin remodeling complexes and target them to specific genomic loci to alter DNA methylation or histone status. These findings suggest an emerging theme of nc RNAs in epigenetic regulation. In this review, we discuss the wide spectrum of nc RNAs in the regulation of DNA methylation and chromatin state, as well as the key questions that needs to be investigated and acknowledging the elegant design of these intriguing macromolecules.展开更多
文摘Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.
文摘Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retention signal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptide into ER, while ER retention signal can make the protein retained in the ER and its derivative protein body. ELISA test indicated that the accumulation level of foreign CpTI protein in sck transgenic tobacco (Nicotiana tabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four times higher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to the increased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be used to increase the accumulation level of foreign protein in transgenic plants and can be widely applied to other related research field in plant genetic engineering.
文摘The vectors carrying the genes coding for the proteins of interest are of unpredictable efficiency in transgenic animals. The expression vector of mammary gland (pINGG) containing GCSF genomic DNA was injected into mouse mammary gland, and expression was detected in the milk of mice. The result showed that mammary gland injection method could provide a convenient transient system to confirm vector validity.
基金Supported by the National Natural Science Foundation of China(30160086,81260164)Key Technology Research and Development Program of Guizhou Province[(2012)3086]+1 种基金the Sixth Group of Scientific and Technological Innovation Talents[(2013)93]Science and Technology Fund of Guizhou[(2011)41]~~
文摘With the development of plant genetic engineering techniques, numerous genetically modified plants have been generated. At the same time, the technologies for detecting transgenic organisms get improved constantly, which also promotes the scientific identification, evaluation and commercial cultivation of transgenic plants. In this review, we evaluate various detection methods for transgenic plants at the level of protein expression.
基金NSFC foundation,Guangdong Province and China Education Ministry joint production-education-research funding Program ( No. 2009B090300198)the Fundamental Research Funds for the Central Universities,HUST( No. M2009060)PhD dissertation Foundation of Huazhong University of Science & Technology
文摘The promoter is a cis-acting element in regulating gene expression. A promoterless plasmid containing UidA gene was transformed into tritordeum by barmbadment. Histochemical analysis of various tissues in transgenic tritordeum was carried to examine tissue-specific expression of GUS(beta-glucuronidase) activity. The pollen-specific promoter was trapped and identified successfully in a transformant line. PCR(polymerase chain reaction) method was used to isolate this pollen-specific promoter. By sequencing and analyzing the amplified fragment from PCR, a part of UidA gene and a flanking sequence were obtained. Some essential elements of plant promoters were found in the sequence. To determine the function of it, the cloned fragment was fused with UidA gene, then cloned and transformed into Triticum durum. The transgenic plant transformed by this vector showed GUS expression only in pollen. Therefore a pollen-specific promoter was isolated successfully.
基金supported by the School of Molecular Cell Biology at the University of Illinois at Urbana-Champaign
文摘Cells are crowded microenvironments filled with macromolecules undergoing constant phys- ical and chemical interactions. The physicochemical makeup of the cells aff)cts various cellular responses, determines cell-cell interactions and influences cell decisions. Chemical and physical properties diff)r between cells and within cells. Moreover, these properties are subject to dynamic changes in response to environmental signals, which often demand adjustments in the chemical or physical states of intracellular molecules. Indeed, cellular responses such as gene expression rely on the faithful relay of information from the outside to the inside of the cell, a process terrned signal transduction. The signal often traverses a complex path across subcellular spaces with variable physical chemistry, sometimes even influencing it. Understanding the molecular states of such signaling molecules and their intracellular environments is vital to our understanding of the cell. Exploring such intricate spaces is possible today largely because of experimental and theoretical tools. Here, we focus on one tool that is commonly used in chemical physics studies light. We summarize recent work which uses light to both visualize the cellular environment and also control intracel- lular processes along the axis of signal transduction. We highlight recent accomplishments in optical microscopy and optogenetics, an emerging experimental strategy which utilizes light to control the molecular processes in live cells. We believe that optogenetics lends un- precedented spatiotemporal precision to the manipulation of physicochemical properties in biological contexts. We hope to use this work to demonstrate new opportunities for chemical physicists who are interested in pursuing biological and biomedical questions.
文摘Recent deep sequencing surveys of mammalian genomes have unexpectedly revealed pervasive and complex transcription and identified tens of thousands of RNA transcripts that do not code for proteins. These non-coding RNAs(nc RNAs) highlight the central role of RNA in gene regulation. nc RNAs are arbitrarily divided into two main groups: The first includes small RNAs, such as mi RNAs, pi RNAs, and endogenous si RNAs, that usually range from 20 to 30 nt, while the second group includes long non-coding RNAs(lnc RNAs), which are typically more than 200 nt in length. These nc RNAs were initially thought to merely regulate gene expression at the post-transcriptional level, but recent studies have indicated that nc RNAs, especially lnc RNAs, are extensively associated with diverse chromatin remodeling complexes and target them to specific genomic loci to alter DNA methylation or histone status. These findings suggest an emerging theme of nc RNAs in epigenetic regulation. In this review, we discuss the wide spectrum of nc RNAs in the regulation of DNA methylation and chromatin state, as well as the key questions that needs to be investigated and acknowledging the elegant design of these intriguing macromolecules.