Plant biotechnology is rife with new advances in transformation and genome engineering techniques. A common requirement for delivery and coordinated expression in plant cells, however, places the design and assembly of transformation constructs at a crucial juncture as desired reagent suites grow more complex. Modular cloning principles have simplified some aspects of vector design, yet many important components remain unavailable or poorly adapted for rapid implementation in biotechnology research. Here, we describe a universal Golden Gate cloning toolkit for vector construction. The toolkit chassis is compatible with the widely accepted Phytobrick standard for genetic parts, and supports assembly of arbitrarily complex T-DNAs through improved capacity, positional flexibility, and extensibility in comparison to extant kits. We also provision a substantial library of newly adapted Phytobricks, including regulatory elements for monocot and dicot gene expression, and coding sequences for genes of interest such as reporters, developmental regulators, and site-specific recombinases. Finally, we use a series of dual-luciferase assays to measure contributions to expression from promoters, terminators, and from cross-cassette interactions attributable to enhancer elements in certain promoters. Taken together, these publicly available cloning resources can greatly accelerate the testing and deployment of new tools for plant engineering.

As of July 17th, 2019 the new Gateway vector website has launched. We are still providing the same Gateway plasmids to the research community but with a more up-to-date look and streamlined ordering process. We are also making available a number of Golden Gate vectors derived from the GreenGate system. Please bear with us as we continue to update the collection and associated information. If you see a problem, or need additional information, please contact us.

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Gateway technology by Invitrogen provides a quick method for cloning a DNA fragment into multiple expression vectors. At the VIB-Ugent Center for Plant Systems Biology, we have constructed over 200 versatile vectors for gene functional analysis in plants and other species. The plasmid accessions offered by PSB comprise Gateway entry clones, intermediary plasmids and destination vectors for transgene construction. Additionally, a growing collection of Golden Gate vectors has been made available as well. This collection contains a wide range of entry modules (promoters, tags and other building blocks) and destination vectors. All plasmids are documented with a graphical map and the corresponding genbank (.gb) file.

Most of the binary constructs we built for Agrobacterium plant transformation are based on the pPZP200 backbone. These constructs carry one of the three most frequently used plant marker genes, selectable with kanamycin, hygromycin or phosphinotricin (Basta). These genes were cloned toward the left border of the T-DNA.

We constructed a novel autonomously replicating gene expression shuttle vector, with the aim of developing a system for transiently expressing proteins at levels useful for commercial production of vaccines and other proteins in plants. The vector, pRIC, is based on the mild strain of the geminivirus Bean yellow dwarf virus (BeYDV-m) and is replicationally released into plant cells from a recombinant Agrobacterium tumefaciens Ti plasmid. pRIC differs from most other geminivirus-based vectors in that the BeYDV replication-associated elements were included in cis rather than from a co-transfected plasmid, while the BeYDV capsid protein (CP) and movement protein (MP) genes were replaced by an antigen encoding transgene expression cassette derived from the non-replicating A. tumefaciens vector, pTRAc. We tested vector efficacy in Nicotiana benthamiana by comparing transient cytoplasmic expression between pRIC and pTRAc constructs encoding either enhanced green fluorescent protein (EGFP) or the subunit vaccine antigens, human papillomavirus subtype 16 (HPV-16) major CP L1 and human immunodeficiency virus subtype C p24 antigen. The pRIC constructs were amplified in planta by up to two orders of magnitude by replication, while 50% more HPV-16 L1 and three- to seven-fold more EGFP and HIV-1 p24 were expressed from pRIC than from pTRAc. Vector replication was shown to be correlated with increased protein expression. We anticipate that this new high-yielding plant expression vector will contribute towards the development of a viable plant production platform for vaccine candidates and other pharmaceuticals.

Dive deep into the fascinating subject of plant vectors in Microbiology, a crucial concept with significant implications in scientific research. Get to grips with the meaning of this term, learn about their key characteristics, and appreciate their essential role in microbiological studies. From understanding different types to exploring practical examples, this resource offers a comprehensive insight into the world of plant vectors. You'll also gain an in-depth understanding about plant viral vectors and their role in plant infection, and the Ti Plasmid Vector for plant transformation. Open up a new world of knowledge through this multifaceted exploration of plant vectors.

For instance, Agrobacterium tumefaciens, a bacterium, is one of the most commonly used vectors for plant genetic engineering. Being itself a plant pathogen, it has the inherent ability to transfer part of its DNA to the plant cell, resulting in a tumour-like growth known as a 'crown gall'. Scienctists ingeniously utilise this mechanism to instead transfer their desired genes into the plant cell.

Plant vectors play a central role in the genetic manipulation of plants, enabling advancements in the fields of molecular biology and plant biotechnology. Through the use of these vectors, scientists can alter the genetic makeup of plants for a host of beneficial purposes such as enhancing resistance to pests, diseases, and environmental conditions, as well as vascular development and flower colour formation.

Moreover, plant vectors also offer a remarkable scope to produce rare compounds which could be precious for pharmaceutical, cosmetic, or food industries. These could range anywhere from the production of insulin to the growth of non-allergenic nuts. The infinite possibilities they present, make them stand at the heart of modern microbiology.

When it comes to the vast and intricate landscape of microbiology, it's crucial to understand that not all plant vectors are the same. Each type has special characteristics that set it apart and make it useful for unique applications.

Plant vectors can be classified based on their origin, the host range they affect, and the methodology they use for gene transfer. Here's a dive into the broad strokes of different types of plant vectors.

Within the realm of microbiology, there are numerous plant vector examples that are employed for various purposes ranging from the study of gene functions to creation of genetically engineered crops. Let's highlight a few salient ones to understand their unique traits and respective roles.

Each plant vector provides a particular way of achieving gene transfer, determined by their biological specificities. This section will delve into a few prototypical examples to delve into their functionalities.

The Agrobacterium tumefaciens, a gram-negative soil bacterium, is arguably the most instrumentally used plant vector. This is primarily because it naturally infects a variety of plant species and introduces its plasmid DNA into the plant cell. Its Ti (Tumour-inducing) plasmid, which causes crown gall disease, has been manipulated to create 'disarmed' vectors - deprived of the disease-causing genes but retaining the gene transfer mechanism.

Rhizobium, another soil bacterium, also effectively transfers genes. However, it is specifically used for legumes as it has a symbiotic relationship with them, helping the host plant fix nitrogen.

Following another route, some vectors are derived from viruses such as Cauliflower Mosaic Virus (CaMV) and Tobacco Mosaic Virus (TMV). These viruses naturally infect plants, insert their genetic material, and make the host produce more of the virus. Scientists can replace a portion of the viral genome with foreign DNA, which the infected plant will then express.

Perhaps one of the most far-reaching applications of plant vectors has been in shaping the 'Golden Rice' project. Here, sequences for two genes involved in the synthesis of beta-carotene (pro-Vitamin A) that are ordinarily unexpressed in the rice endosperm were introduced using Agrobacterium. The resultant genetically modified (GM) rice has the potential to alleviate vitamin A deficiency among populations relying heavily on rice.

Plant vectors also made it possible to generate 'Frostban' strawberries that can tolerate frost better. To achieve this, scientists introduced a gene coding for an antifreeze protein from the flounder fish into strawberry plants.

A fascinating intersection of plant biology and microbiology occurs in the form of plant viral vectors. These are viruses specially designed to carry and transfer genetic material into plant cells, a concept hinging on the natural ability of viruses to infiltrate host cells and integrate their genome into the host's. This trait, which makes viruses pathogenic in nature, has been harnessed for beneficial purposes in plant genetic engineering and functional genomics. 0852c4b9a8

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