Chloroplast Genetic Engineering in Higher Plants

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.

Advances in Cold Tolerance Genes and Their Application in Genetic Engineering of Plant for Cold Tolerance

Low temperature is one of the main environmental stress factors influenc- ing plant growth and development and crop yield. Cold tolerance genes and progress of their application in genetic engineering of plant for cold tolerance were reviewed comprehensively and systematically from the aspect of genes that are in- volved in biosynthesis of osmotic substances, genes coding fatty acid desaturation enzymes, antifreeze protein genes, genes coding antioxidant enzymes and so on, aiming at laying the foundation for genetic improvement of cold tolerance and breeding of plants.

Crop Resources Ethic in Plant Genetic Engineering and Fortune Transfer Between Generations

The relation between human and crop resources belongs to the ethic of resources exploitation. The purposes of discussing the ethic of crop resources are to protect the ecology and safety of crops, to gain sustainable development, furthermore, to choose and form the production structure that is favorable to saving crop resources and protecting the ecology of crops. Plant genetic engineering is the technology of molecule breeding of rearrangement of inheritance materials at the level of molecule directionally, of improving plant properties and of breeding high quality and yield varieties of crops. The prominent effects of the technology on the crop ecological system are human subjective factors increasing as well as violating the nature and intensifying the conflict between human being and nature. Therefore, in plant genetic engineering, crop resources exploitation should follow certain ethic principles. Under the theory of ethics of natural resources, by the means of biologinal-statistics, the author systematically analyzed the possible model of crop resources transfer between generations as well as the transfer mode of magnitude of real materials and magnitude of value.

Conifer genetic engineering: Particle bombardment and Agrobacterium-mediated gene transfer and its application in future forests

Many important advances in forest biotechnology have been made. The use of genetic transformation and the ap-plications of transgenic trees in modern forestry is now an important field. Two basic methodologies particle bombardment and Agrobacterium-mediated transformation have been used on conifers. However, routine procedures exist for only a limited number of conifers. As a result only a few species have been successfully transformed into stable transgenic plants. The use of a particle bombardment has been more successful and transgenic plants have been produced in Picea abies, Picea glauca, Picea mariana, and Pinus radiata, although the level of production of stable transgenic plants is lower than that of Agrobacte-rium. At present, breeding programs have been directed toward improving bole shape, growth rate, wood properties, and quality, as well as toward improving root and shoot performance, pest resistance, stress tolerance, herbicide resistance, and ability to resist stresses, which will drive forestry to enter a new era of productivity and quality. This article provides a brief overview of the current state of knowledge on genetic transformation in conifers.

Genetic engineering and lignin biosynthetic regulation in forest tree species

Genetic engineering of forest tree species is regarded as a strategy to reduce worldwide pressure on natural forests, to conserve genetic resources and ameliorate stress on global climate, and to meet growing demand for forest wood and timber products. Genetic engineering approaches toward the control or management of fungal pathogens, arthropod herbivores, bacterial and viral diseases, the use of pest resistance genes, and weed competitors are being studied. Although the production of transgenic trees is relatively recent and only a few species have been successfully genetically engineered in forest tree species, very useful and valuable information is available on the application of transgenic trees. Genes involved in important agricultural traits such as herbicide resistance, insect resistance, and wood quality have been isolated and have been used to genetically engineer trees. New technologies of plant molecular biology and genomics now make it possible high-efficient genetic improvement of forest trees. Genetic engineering promises to expand greatly the potential for genetic manipulation as new genes of commercial interest are discovered and utilized. Lignification is a process essential to the nature and evolution of vascular plants that is still poorly understood, even though it has been studied for more than a century. Recent studies on mutant and transgenic plants indicate that lignification may be far more flexible than previously realized. Rines with a mutation affecting the biosynthesis of the major lignin precursor, coniferyl alcohol, show a high level of an unusual subunit, dihydroconiferyl alcohol. It is also unusual as a plant polymer in that there are no plant enzymes for its degradation. These results have significant implications regarding the tradiational definition of lignin, and highlight the need for a better understanding of the lignin precursor biosynthetic pathway. In this review, we describe the progress made recently in genetic engineering of forest tree species.

A Review on Creating Male Sterility in Vegetable Crops by Genetic Engineering

With the deep researches on male sterility genetic engineering of plants, several strategies creating male sterile materials have been developed, such as causing pollen abortion by cytotoxic genes, antisense RNA or RNAi silencing the expression of genes related to pollen development, early degradation of tapetum callose leading to male sterility. Male sterile transgenic plants can be obtained through genetic transformation with related genes destroying or interfering with pollen or anther development. Male sterile cauliflower, tomato, cabbage, etc. have been developed in this way, and some begin to be used to produce hybrid seed. Appling some techniques can also maintain and restore the male sterility. These related researches will effectively promote the heterosis utilization and the development of crop breeding. This paper mainly presents their principles and applications in vegetable crops.

Osmoregulation Mechanism of Drought Stress and Genetic Engineering Strategies for Improving Drought Resistance in Plants

Drought, one of the main adverse environmental factors, obviously affected plant growth and development. Many adaptive strategies have been developed in plants for coping with drought or water stress, among which osmoregulation is one of the important factors of plant drought tolerance. Many substances play important roles in plant osmoregulation for drought resistance, including proline, glycine betaine, Lea proteins and soluble sugars such as levan, trehalose, sucrose, etc. The osmoregulation mechanism and the genetic engineering of plant drought-tolerance are reviewed in this paper.

Recent advances in the molecular genetics of resin biosynthesis and genetic engineering strategies to improve defenses in conifers

Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a monoterpene synthase, a sesquiterpene synthase, and a diterpene synthase. Tree killing bark beetles and their vectored fungal pathogens are the most destructive agents of conifer forests worldwide. Conifers defend against attack by the constitutive and inducible production of oleoresin that accumulates at the wound site to kill invaders and both flush and seal the injury. Although toxic to the bark beetle and fungal pathogen, oleoresin also plays a central role in the chemical ecology of these boring insects. Recent advances in the molecular genetics of terpenoid biosynthesis provide evidence for the evolutionary origins of oleoresin and permit consideration of genetic engineering strategies to improve conifer defenses as a component of modern forest biotechnology. This review described enzymes of resin biosynthesis, structural feathers of genes genomic intron and exon organization, pathway organization and evolution, resin production and accumulation, interactions between conifer and bark beetle, and engineering strategies to improve conifer defenses.

Preparation of Conotoxin MrVIB by Genetic Engineering Technology

[ Objective] The disulfide-rich conotoxin MrV1B was produced by simple and fast genetic engineering method, to find new efficient ways for the synthesis of natural active conotoxins. [Method] Primers of conotoxin gene MrVIB were synthesized to construct expression vectors pET22b（ ＋ ）/His-Xa-MrVIB and pET32a/Trx-EK-MrV1B, which were transformed into BL21 （DE3）pLysS and expressed under induction by IPTG. Recombinant proteins were purified by affinity chromatography using Ni-NTA agarose column, and the expression of the recombinant proteins was analyzed by Tricine-SDS-PAGE electrophoresis. [ Result] The recombinant conotoxins His-Xa-MrVIB and Trx-EK-MrVIB were effectively expressed in E. coli, and purified by one-step affinity chromatography, and the purity of the recombinant conotoxins was greater than 90%. [ Conclusion] The conotoxin MrVIB was effectively secreted and expressed by genetic engineering method, which could solve the problems in chemical synthesis of conotoxins including low yield, high cost and difficult purification.

Advantages and Disadvantages of Transgenic Animal Technology with Genetic Engineering

Transgenic animal technology has been one of the fastest growing biotechnology areas. The exogenous genes have been introduced into the animal genome by genetic engineering, so that these genes can be inherited and expressed by offspring to produce desired traits or evaluate function in elite livestock breeds. There are several methodologies for the production of transgenic animals, i.e., （1） microinjection of genes into pronuclei of fertilized ova; （2） DNA transfer by retroviruses; （3） injection of embryonic germ （EG）/embryonic stem （ES） cells previously treated with foreign DNA; （4） DNA transfer into cells and embryos with using liposomes; （5） exogenous DNA transfer while in vitro fertilization by using sperm; （6） electroporation of DNA into sperm, embryos or ova; （7） biolistics; （8） nuclear transfer （NT） with somatic cells, EG or ES cells; （9） germ line stem cell-mediated; （10） gene targeting; （! 1） gene silencing technology with RNA interference; （12） induced pluripotent stem cell; （13） zinc-finger nuclease gene targeting technology. Gene farming is one of the newest and most promising areas in modern biotechnology. Cattle, goats, sheep, pigs and rabbits are the main farm livestock species and fish is also used in transgenic technology. The question of ＂why make transgenic animals？＂ is very important. Some of the answers to this question are： （1） to obtain new knowledge; （2） to solve the genetic code; （3） to create genetic disease models; （4） to study the genetic control of physiological systems; （5） to improve animal production traits; （6） to produce new animal products. Transgenic technology is one of the main and important tools in the finding solutions to problems of growing population with their applications to different organisms, and takes more attention and interest every day. Transgenic technology creates opportunities and areas to play with organisms to fulfill the demands of people. Because of this, this paper based on mainly transgenic applications to take people＇s attention and exhibit its importance.

Editor's Choice——Application of genetic engineering for the treatment of neurodegenerative diseases

Gene therapy has been shown to be an effective method for protecting neural functions in the substantia nigra,

Approaches to Detecting Gene-Environment Interactions in Human Variation Using Genetic Engineering, Remote Sensing and GIS

Elucidation of the relationships between genetic polymorphisms and environmental exposures can provide insights into the pathways and mechanisms underlying complex traits. A new approach was used to detect G×E （gene-environment） interactions involved in human skin pigmentation variation to better understand the adaptive evolution of skin pigmentation. Specifically, we used genetic engineering, remote UVR （ultraviolet radiation） sensing and GIS （geographic information systems） to integrate the analysis of genetic and environmental factors into a coherent biological framework. Since we expected to generate large datasets for this multidimensional analysis, we used PCA （principal components analysis） as a spatial statistical analysis technique for analyzing the G×E interactions. The results suggest that skin pigmentation may be affected by mutations induced by UVR and support the hypothesis that global variation in skin pigmentation may be the result of localized adaptation to different UVR conditions via natural selection. Analyzing the relationships between heterozygous frequencies for SNP （single nucleotide polymorphism） loci and seasonal UVR levels as the environment changes will help elucidate the selective mechanisms involved in the UVR-induced evolution of skin pigmentation. Skin pigmentation fulfills the criteria for a successful evolutionary G×E interactions model.

Development Prospects of Genetic Engineering Pharmaceuticals

Human science and technology continue to advance over time.In the future,universal drugs will gradually fade out of our lives with the accumulation of time.With the advancement of genetic engineering,future genetic engineering drugs will be based on each difference and due to It differs from person to person,and the development of genetic engineering pharmaceuticals will make breakthroughs.

Human a Type Genetic Engineering Interference Essence Injection

The highest-level interference essence against virus and turnour genetic engineering medicine is a new type created in the 1980s. Compared with chemical medicines, the interference essence has a special effect in the treatment of viruses and tumours. The human a, type genetic engineering interference essense is prepared by the Institute of Viruses of the Chinese Academy of Preventive Medical Sciences, the Shanghai Vaccine

Eradication of malaria through genetic engineering:the current situation

Malaria is an intra-cellular parasitic protozoon responsible for millions of deaths annually.Host and parasite genetic factors are crucial in affecting susceptibility to malaria and progression of the disease.Recent increased deployment of vector controls and new artemisinin combination therapies have dramatically reduced the mortality and morbidity of malaria worldwide.However, the gradual emergence of parasite and mosquito resistance has raised alarm regarding the effectiveness of current artemisinin-based therapies.In this review,mechanisms of anti-malarial drug resistance in the Plasmodium parasite and new genetically engineered tools of research priorities are discussed.The complexity of the parasite lifecycle demands novel interventions to achieve global eradication.However,turning laboratory discovered transgenic interventions into functional products entails multiple experimental phases in addition to ethical and safety hurdles.Uncertainty over the regulatory status and public acceptance further discourage the implementation of genetically modified organisms.

Advances in homology directed genetic engineering of human pluripotent and adult stem cells

The ability to introduce precise genomic modifications in human cells has profound implications for both basic and applied research in stem cells, ranging from identification of genes regulating stem cell self-renewal and multilineage differentiation to therapeutic gene correction and creation of in vitro models of human diseases. However, the overall efficiency of this process is challenged by several factors including inefficient gene delivery into stem cells and low rates of homology directed site-specific targeting. Recent studies report the development of novel techniques to improve gene targeting efficiencies in human stem cells; these methods include molecular engineering of viral vectors to efficiently deliver episomal genetic sequences that can participate in homology directed targeting, as well as the design of synthetic proteins that can introduce double-stranded breaks in DNA to initiate such recombination events. This review focuses on the potential of these new technologies to precisely alter the human stem cell genome and also highlights the possibilities offered by the combination of these complementary strategies.

Efficient and Soluble Expression of α Protein of Clostridium perfringens and Preparation of Genetic Engineering Subunit Vaccine

[Objective] The paper was to develop genetic engineering vaccine that can express α exotoxin antigen protein efficiently without destroying its immunogenicity for preventing and controlling the diseases caused by Clostridium perfringens. [Method] Efficiently expressed soluble recombinant α protein was obtained from Escherichia coli expression system by optimizing codon,removing signal peptide,selecting sequences with better hydrophilicity and antigenicity,and optimizing expression conditions. [Result] Mice obtained higher serum antibody level when immunized by α protein,and the immune protection rates against type A,type B,type C and type D C. perfringens were 100%,90%,85% and 90%,respectively. The antibody titer of mice within 7-14 d after the third immunization reached the peak. [Conclusion]The α protein has good immunogenicity,and can be further used to develop genetic engineering subunit vaccines for preventing C. perfringens.

Genetic Engineering of Field, Industrial and Pharmaceutical Crops

Ability to modify plants at the genomic level by advanced molecular technology has enhanced the scope of improvements in plant traits attempted earlier through conventional breeding methods. Techniques such as genetic transformation have opened new vistas whereby functional genes, not commonly present in a particular species can be added from other species. The traits incorporated into the genetically engineered plants in the beginning were confined to those governed by dominant genes, e.g. insecticide resistance and herbicide tolerance but advancements with time now also permit the transfer of complexly inherited traits such as drought and cold tolerance. Transgenic technology is also useful in understanding gene expression and metabolic pathways which can then be used to harness the full genomic potential of the plant. This review presents a narrative on development of transgenics and their use for the improvement of field, industrial and pharmaceuticals crops. In addition, discussions are made on current status on genetically modified crops, hurdles to genetic engineering, overcoming strategies and future scope.

Construction of Multi-Specific Antibody by Genetic Engineering and Its Progress in Tumor Therapy

Targeted treatment of cancer with monoclonal antibodies increases the benefit for patients. In order to improve the anti-tumor activity of monoclonal antibodies, multi-specific antibodies have entered the research field. The emergence of various techniques to produce multi-specific recombinant antibody molecules has led to the selection of target combinations in various forms. To date, only a few multi-specific constructs have entered phase III clinical trials, in contrast to classical monoclonal antibodies. Some of the format options are outlined from a technical point of view. We focus on the achievements and prospects of the underlying technologies for generating biand multispecific antibodies.