여기에 제시된 연구는 찰스 강 (Charles River)에 의해 지원되었다.

<ol>
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저자는 찰스 강 (Charles River)의 직원입니다.

이 프로토콜은 연구원의 시설에서 제공 교육 실습을 보완하기위한 동물 처리 및 약물 관리에 대한 소개로 볼 수 있습니다. 사용되며 물질 관리의 노선이 연구 프로토콜이나 윤리위원회 프로토콜이 작성 될 때 실험 설계에 고려하고해야 덮개의 의미합니다. 동물 관련 절차에 교육 연구의 성공에 매우 중요합니다. 대부분의 실험을 수행하려면, 동물 연구 직원이 처리해야하며, 더 나은 동물 취급, 적은 동물 3 강조했다. 부드러운 인간의 연락처로 동물을 Accustoming은 스트레스를 줄이고 동물보다 다루기 쉬운 연구 과목 4,5 할 수 있습니다. 취급 스트레스는 연구 6 몇 가지 유형에 영향을 표시하고이뿐만 아니라 다른 사람에 영향을 미칠 수 있습니다 수 있습니다되었습니다. 설치류의 규제는 (예정주의를 달성하지만, 회사 처리해야그립은 쥐와 핸들러에 부상을 초래할 가능성이)와 실제 짧은 기간 동안해야합니다. 구속 방법은 일반적으로 동물의 크기에 따라 선택하거나 접근을 모색하고 있습니다. 예를 들어, 가능하지만, 안녕하세요,이 성인 쥐를 취급,​​ 자주 처리기가 미숙 특히 경우, 쥐에 의해 강한 저항을 만났 있습니다. 손으로 마우스 또는 쥐를 개최하는 것은 꼬리 정맥에 액세스 어렵고 구속 장치가 자주 여전히 가능한 동물을 유지하기로 선택되어 있는지 확인 할 수 있습니다. 연구자들이 동물을 처리 할 때, 그들은 종종 더 연구를위한 화합물이나 생물학을 관리하기 위해 찾고 있습니다. 물질 관리의 경로는 특정 실험을 흡수, 생체 이용률, 그리고 적합성에 영향을 미칠 수 있습니다. 다양한 노선과 친숙 함은 연구 최선의 방법으로 자신의 물질을 관리 할 수​​있는 능력을 가진 연구자를 제공해야합니다. 물질의 빠른 흡수를 촉진 예를 들어, 경로연구자가 느린 - 연기 방식으로 물질을 관리하기를 원하는 경우, 이러한 정맥 주사 또는 intraperitoneal으로 사용할 수 없습니다. 볼륨, 장비 및 용질에 대해 이러한 기술과 고려 사항 중 일부 최근 리뷰가 터너 외하여 두 논문에서 찾을 수 있습니다. 1,7는 물질이 실험실 동물에 투여 할 때마다, 고려는 장비와 물질 (표 1에서 설명)의 볼륨의 적절한 크기로 제공해야합니다. 잘못 크기의 장비 나 대형 볼륨 불편, 부상, 또는 동물의 죽음에 발생할 수 있습니다. 일반적으로, parenterally 실시 물질 연구 목표 (즉, 세균 연구)이 불가능 할 곳을 제외하고 무균 수 있습니다. 화합물 및 생체은 동물에 최소 영향을 미칩니다 용질 또는 차량에 있어야합니다. physiologic 산도 (7.3 -7.4)은 특히 피하, 근육, 그리고 intrape를 들어, 일반적으로 잘 인정합니다ritoneal 노선. 이 노선에서 관리 물질의 비 physiologic pH의 수준은 통증이나 괴사 및 조직 손상 될 수 있습니다. 산도의 넓은 범위는 intragastric와 정맥 주사 경로를 7 용납되어 있습니다. 작은 설치류에서 또 다른 중요한 고려 사항은 룸 온도 유체의 대량이 주어진 경우 냉각의 가능성입니다. 유체 특히 병 동물의 지원에 intravenously 또는 intraperitoneally 관리되는 경우는 체온 (37 ° C)로 예열해야합니다. 이 프로토콜에서 설명하는 관리의 노선 마스터 간단하고, 일반적으로 마취를 필요로하지 않는, 일반적으로 많은 연구 프로그램에 사용되는 것들입니다. 행정의 노선의 거의 무한한 다양성 intracranial, intrathecal, 경막 외, intratracheal, intraosseous, 그리고 이름 intraarticular하지만 몇을 포함하지만, 가능합니다. 행정 이러한 전문 노선의 교육 사람들 대체에서 추구되어야한다O는 경로 좋은 결과와 다양한 경험을 보유하고 있습니다. intratracheal 기지보다 방법, 동물에서 비강 경로는 일반적으로 더 &quot;자연&quot;을 통해 폐에 도입 물질을 연구하는 데 사용됩니다. 마우스 및 쥐도 의식 동물에 어렵지 않습니다 그래서 그들이 유체의 매우 작은 양을 흡입하는 유도, 고맙게 여기게 코 - breathers 있습니다. 코 점막은 혈관, 일부 물질의 비강 관리를 잘 공급되어 있기 때문에 정맥 주사 관리와 유사 할 수 있습니다. 그 흡수를 손상 할 수 있으므로이 경로는하지만, 비염과 동물에 권장하지 않습니다. 비강 경로에서 큰 볼륨을 관리하려고 시도 dyspnea 또는 동물의 구조 될 수 있습니다. 근육 주사는 물질의 빠른 흡수를 제공합니다. 근육 주사는 쥐와 작은 크기와 상응 작은 근육으로 인해 마우스에 도전 할 수 있습니다. 그들은 뒷다리로 진행됩니다s입니다. 좌골 신경의 손상의 가능성으로 인해, quadriceps femoris의 선택의 근육이다. 피하와 intradermal 노선 모두 피부를 포함하지만, subcutis 대 피부에 배치 물질의 생체 이용률의 차이가 있습니다. 피하 관리는 종종 이러한 정맥 주사 또는 intraperitoneal 등 다른 노선보다 느린 흡수와 함께 &quot;증착&quot;루트로 간주됩니다. Intradermal 관리는 일반적으로 물질 등 도움이되는 - 항원 혼합물로 immunostimulatory 물질의 아주 작은 볼륨에 사용됩니다. 두 경우 모두, 관리 물질 physiologic pH의 비 자극을해야합니다. 이 쥐을 억제를위한 일반적으로 사용되는 사이트이기 때문에 Intradermal 또는 피하 주사는, 안녕하세요,에 수행 할 수 없습니다. 정맥 주사와 intraperitoneal 관리는 종종 동물의 상응하는 것으로 간주됩니다. 주입의 정맥 주사 경로를 더 연구를 제공합니다화합물의 apid 이해하지만, intraperitoneal 관리가 구강 관리 8 대략적으로 비슷 고려되어야하는 동안. 부적절한 버퍼 경우는 통증의 원인이 될 수 있습니다으로 케어 화합물 관리 intraperitoneally로 촬영해야합니다. 설치류의 정맥 bolus 관리의 일반적인 경로는 꼬리 정맥을 통해 수 있습니다. 물질의 만성 정맥 주사 관리가 필요하다면, 정맥 또는 동맥 캐 뉼러의 주입이 고려되어야한다. intravenously 관리 물질 무균 전달해야하며 intravenously 관리하는 안전 표시 될 것입니다. 예를 들어, 용혈, 혈전증, 또는 vasculitis을 유도 할 수 있습니다 물질 정맥 주사 관리에 적합하지 않습니다. intragastric 또는 구두 gavage 경로는 종종 인간의 일반적인 주입 경로를 모방하는 데 사용됩니다. 음식이나 물을 통해 구두 관리에 비해 또한 물질의 정확한 주입 할 수 있습니다. 화합물의 생체 이용율gavage를 통해 관리 동물의 먹이 / 공복 상태뿐만 아니라 화합물 또는 생물학적의 용질 또는 차량에 따라 달라집니다. Gavage 또는 수유 바늘이 사용되는 동물에 대한 적절한 크기이어야하고, 일회용 gavage 바늘이 실제하지 않은 경우, 동물 사이에 청소해야합니다. gavage로 인한 부상은 드문 일이 아니에요 있으며, 폐 또는 위 또는 식도의 파열에 물질의 증착이 포함되어 있습니다. 교육은 경험이 풍부한 자에 의해 감독과 깨어있는 동물 gavage 전에 anesthetized 동물 (euthanized 될 것이다)를 먼저 euthanized 동물에 수행되어야한다이 시도됩니다. 깨어있는 동물 최초의 gavage 시도는 프로 시저가 틀려서되면 부상을 입을되지 않는 생리 등의 평균 크기의 동물과 물질의 작은 볼륨을, 포함한다. 동물 밀접하게 gavage와 euthanized 필요한 경우 이후, 파랑, 출혈, 또는 초과 타액의 분비를 돌려 같은 웃음과 같은 고통의 흔적에 대한 평가해야합니다. 만약 euthanasia은 동물이 gavage 절차가 실패 이유를 확인 necropsied해야 필요합니다.

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 코멘트 </td></tr><tr><td> 바늘 </td><td> 여러 </td><td> 여러 </td><td> 바늘이 게이지 및 길이 모두에서 판매하고 있습니다. 주문하기 전에 모두 확인하십시오. </td></tr><tr><td> 주사기 </td><td> 여러 </td><td> 여러 </td><td> 항상 관리 할 볼륨에 대해 적절한 크기를 선택합니다. </td></tr><tr><td> DecapiCones </td><td> Braintree 과학 </td><td> DC-200, DCL-120, MDC-200 </td><td> 마우스 및 쥐 크기 사용할 수 있습니다. </td></tr><tr><td> 쥐 restrainer </td><td> 하버드 장치, Braintree 과학, Plas-랩, 다른 </td><td></td><td> 맑은 Plexiglas, 조정 플라스틱, 및 마우스 및 쥐에 대한 크기에서 사용할 수 있습니다. </td></tr><tr><td> 50 ML 원뿔 관 </td><td> 여러 </td><td> 및주의SP; </td><td></td></tr><tr><td> 수유 바늘 </td><td> VWR, Popper와 아들 </td><td> 여러 </td><td> 바늘 게이지하고 위에 설명 된대로 동물 길이를 맞추기. 모두 일회용과 재사용 먹이 바늘을 사용할 수 있습니다. </td></tr></tbody></table>

manual restraint and common compound administration routes in mice and rats

안전하고 효과적으로 마우스 및 쥐를 제지 할 수 있다는 것은 연구의 중요한 부분입니다. 마우스 및 쥐와 함께 자신과 편안하게 작업하면 처리 및 규제 방법의 기본 역량이 필요합니다. 이 문서는 안전하게 동물을 처리하는 데 필요한 기본 원칙을 제시합니다. 특별히 고안된 구속 개체 한 손, 두 손, 그리고 구속이 설명됩니다. 종종, 동물의 연구 또는 시험 사용의 또 다른 부분은 마우스 및 쥐에 대한 화합물의 효과적인 관리이다. 가능한 관리 노선의 큰 번호 (만 동물의 크기와 기관에 의해 제한)가 있지만, 대부분은 연구에 정기적으로 사용되지 않습니다. 이 동영상은 정맥 주사, 근육, 피하, 그리고 구두 gavage 등의보다 일반적인 경로, 여러을 설명합니다. 이 문서의 목적은 기본 구속 및 약물 관리 노선에 이러한 기술에 익숙 뷰어를 노출하는 것입니다. 이 동영상은귀하의 시설에서 교육에 - 손 필수를 대체하지 않습니다,하지만 그건 교육을 보강하고 보완하기위한 것입니다.

Watch this Scientific Journal Video about Manual Restraint and Common Compound Administration Routes in Mice and Rats at JoVE.com

Manual Restraint and Common Compound Administration Routes in Mice and Rats

연구 설치류와 함께 안전하고 편안하게 일하는 것은 처리 및 규제 방법의 핵심 역량이 필요합니다. 이 문서는 안전하게 처리하고 효과적으로 마우스 및 쥐에 화합물을 관리하는 데 필요한 기본 원칙을 제시합니다.

수동 규제와 마우스 및 쥐 일반적인 합성 관리 경로

Being able to safely and effectively restrain mice and rats is an important part of conducting research.  Working confidently and humanely with mice and ...

manual-restraint-common-compound-administration-routes-mice

Nanyang Technological University

Research

Education

JoVE Core

JoVE Journal

Pharmacology

Biology

Unit 1

General Pharmacological Principles

SCIENTISTS IN ACTION

289.7K 조회수.  Charles River. 연구 설치류와 함께 안전하고 편안하게 일하는 것은 처리 및 규제 방법의 핵심 역량이 필요합니다. 이 문서는 안전하게 처리하고 효과적으로 마우스 및 쥐에 화합물을 관리하는 데 필요한 기본 원칙을 제시합니다.

비디오: Manual Restraint and Common Compound Administration Routes in Mice and Rats

Induction and Monitoring of Active Delayed Type Hypersensitivity (DTH) in Rats

Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes that infiltrate the site of injection of an antigen against which the immune system has been primed. The inflammatory reaction is characterized by redness and swelling of the site of antigenic challenge. It is a convenient model to determine the in vivo efficacy of immunosuppressants. Cutaneous DTH can be induced either by adoptive transfer of antigen-specific T lymphocytes or by active immunization with an antigen, and subsequent intradermal challenge with the antigen to induce the inflammatory reaction in a given skin area. DTH responses can be induced to various antigens, for example ovalbumin, tuberculin, tetanus toxoid, or keyhole limpet hemocyanin (KLH).
Here we demonstrate how to induce an active DTH reaction in Lewis rats. We will first prepare a water-in-oil emulsion of KLH, our antigen of interest, in complete Freund's adjuvant and inject this emulsion subcutaneously to rats. This will prime the immune system to develop memory T cells directed to KLH. Seven days later we will challenge the rats intradermally on the back with KLH on one side and with ovalbumin, an irrelevant antigen, on the other side. The inflammatory reaction will be visible 16-72 hours later and the red and swollen area will be measured as an indication of DTH severity.

Induction and Monitoring of Active Delayed Type Hypersensitivity DTH in Rats

Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes. Here we demonstrate how to induce active DTH in Lewis rats and monitor the inflammatory response.

쥐에서 Active 지연 유형 과민성 (DTH)의 유도 및 모니터링

Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes that infiltrate the site of injection of ...

연구

생물학

Induction and Monitoring of Adoptive Delayed-Type Hypersensitivity in Rats

Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes that infiltrate the site of injection of an antigen against which the immune system has been primed. The inflammatory reaction is characterized by redness and swelling of the site of antigenic challenge. It is a convenient model to determine the in vivo efficacy of immunosuppressants. Cutaneous DTH can be induced either by adoptive transfer of antigen-specific T lymphocytes or by active immunization with an antigen, and subsequent intradermal challenge with the antigen to induce the inflammatory reaction in a given skin area. DTH responses can be induced to various antigens, for example ovalbumin, tuberculin, tetanus toxoid, or keyhole limpet hemocyanin. Such reactions can also be induced against autoantigen, for example to myelin basic protein (MBP) in rats with experimental autoimmune encephalomyelitis induced with MBP, an animal model for multiple sclerosis (1).
Here we demonstrate how to induce an adoptive DTH reaction in Lewis rats. We will first stimulate ovalbumin-specific T cells in vitro and inject these activated cells intraperitoneally to naive rats. After allowing the cells to equilibrate in vivo for 2 days, we will challenge the rats with ovalbumin in the pinna of one ear, while the other ear wil receive saline. The inflammatory reaction will be visible 3-72 hours later and ear thickness will be measured as an indication of DTH severity.

Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T (TEM) lymphocytes. Here we demonstrate how to activate antigen-specific TEM cells, induce adoptive DTH in Lewis rats and monitor the inflammatory response.

고양이의 입양 지연 유형 과민성의 유도 및 모니터링

Changes in Mammary Gland Morphology and Breast Cancer Risk in Rats

Studies in rodent models of breast cancer show that exposures to dietary/hormonal factors during the in utero and pubertal periods, when the mammary gland undergoes extensive modeling and re-modeling, alter susceptibility to carcinogen-induced mammary tumors. Similar findings have been described in humans: for example, high birthweight increases later risk of developing breast cancer, and dietary intake of soy during childhood decreases breast cancer risk. It is thought that these prenatal and postnatal dietary modifications induce persistent morphological changes in the mammary gland that in turn modify breast cancer risk later in life. These morphological changes likely reflect epigenetic modifications, such as changes in DNA methylation, histones and miRNA expression that then affect gene transcription . In this article we describe how changes in mammary gland morphology can predict mammary cancer risk in rats. Our protocol specifically describes how to dissect and remove the rat abdominal mammary gland and how to prepare mammary gland whole mounts. It also describes how to analyze mammary gland morphology according to three end-points (number of terminal end buds, epithelial elongation and differentiation) and to use the data to predict risk of developing mammary cancer.

Our protocol describes how to dissect the rat abdominal mammary gland and how to prepare mammary gland whole mounts. It also describes how to analyze mammary gland morphology using three end-points (number of terminal end buds, epithelial elongation and differentiation) and to use these results to predict mammary cancer risk in rats which were exposed to dietary modifications in utero or during prepuberty.

쥐에서 유선의 형태론과 유방암 위험의 변화

Studies in rodent models of breast cancer show that exposures to dietary/hormonal factors during the in utero and pubertal periods, when the mammary gland ...

Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal colony. Proper collection of tissues for further histological processing can impact the quality of testing results. This article describes the conduct of a basic gross examination including identification of heart, liver, lungs, kidneys, and spleen, as well as how to collect those organs. Additionally four of the more difficult tissue/sample collection techniques are demonstrated. Lung collection and perfusion can be particularly challenging as the tissue needs to be properly inflated with a fixative in order for inside of the tissue to fix properly and to enable thorough histologic evaluation. This article demonstrates the step by step technique to remove the lung and inflate it with fixative in order to achieve optimal fixation of the tissue within 24 hours. Brain collection can be similarly challenging as the tissue is soft and easily damaged. This article demonstrates the step by step technique to expose and remove the brain from the skull with minimal damage to the tissue. The mesenteric lymph node is a good sample type in which to detect many common infectious agents as enteric viruses persist longer in the lymph node than they are shed in feces. This article demonstrates the step by step procedure for locating and aseptically removing the mesenteric lymph node. Finally, identification of infectious agents of the respiratory tract may be performed by bacterial culture or PCR testing of nasal and/or bronchial fluid aspirates taken at necropsy. This procedure describes obtaining and preparing the respiratory aspirate sample for bacterial culture and PCR testing.

This article describes the procedures for conducting a basic postmortem examination of a mouse or rat, and the collection of basic organs, as well as more challenging sample types from for histological, microbiological, and PCR evaluation.

진단 검시와 고양이와 마우스의 선택 조직 및 샘플 수집

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal ...

Ovariectomy and 17&beta;-estradiol Replacement in Rats and Mice: A Visual Demonstration

Estrogens are a family of female sexual hormones with an exceptionally wide spectrum of effects. When rats and mice are used in estrogen research they are commonly ovariectomized in order to ablate the rapidly cycling hormone production, replacing the 17&beta;-estradiol exogenously. There is, however, lack of consensus regarding how the hormone should be administered to obtain physiological serum concentrations. This is crucial since the 17&beta;-estradiol level/administration method profoundly influences the experimental results1-3. We have in a series of studies characterized the different modes of 17&beta;-estradiol administration, finding that subcutaneous silastic capsules and per-oral nut-cream Nutella are superior to commercially available slow-release pellets (produced by the company Innovative Research of America) and daily injections in terms of producing physiological serum concentrations of 17&beta;-estradiol4-6. Amongst the advantages of the nut-cream method, that previously has been used for buprenorphine administration7, is that when used for estrogen administration it resembles peroral hormone replacement therapy and is non-invasive. The subcutaneous silastic capsules are convenient and produce the most stable serum concentrations. This video article contains step-by-step demonstrations of ovariectomy and 17&beta;-estradiol hormone replacement by silastic capsules and peroral Nutella in rats and mice, followed by a discussion of important aspects of the administration procedures.

Ovariectomy and 17&#946;-estradiol Replacement in Rats and Mice: A Visual Demonstration

Ovariectomy and subsequent replacement of 17&beta;-estradiol by means of silastic capsules and peroral Nutella are demonstrated in rats and mice.

고양이와 쥐의 Ovariectomy 및 17β-estradiol 대체 : 비주얼 데모

Estrogens are a family of female sexual hormones with an exceptionally wide spectrum of effects. When rats and mice are used in estrogen research they are ...

Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR

A unique open reading frame (ORF) Z3276 was identified as a specific genetic marker for E. coli O157:H7. A qPCR assay was developed for detection of E. coli O157:H7 by targeting ORF Z3276. With this assay, we can detect as low as a few copies of the genome of DNA of E. coli O157:H7. The sensitivity and specificity of the assay were confirmed by intensive validation tests with a large number of E. coli O157:H7 strains (n = 369) and non-O157 strains (n = 112). Furthermore, we have combined propidium monoazide (PMA) procedure with the newly developed qPCR protocol for selective detection of live cells from dead cells. Amplification of DNA from PMA-treated dead cells was almost completely inhibited in contrast to virtually unaffected amplification of DNA from PMA-treated live cells. Additionally, the protocol has been modified and adapted to a 96-well plate format for an easy and consistent handling of a large number of samples. This method is expected to have an impact on accurate microbiological and epidemiological monitoring of food safety and environmental source.

Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR

A qPCR assay was developed for detection of Escherichia coli O157:H7 targeting a unique genetic marker, Z3276. The qPCR was combined with propidium monoazide (PMA) treatment for live cell detection. This protocol has been modified and adapted to a 96-well plate format for easy and consistent handling of numerous samples

라이브의 검출<em&gt; 대장균</em&gt; O157 : PMA-qPCR에 의한 H7 셀

A unique open reading frame (ORF) Z3276 was identified as a specific genetic marker for E. coli O157:H7. A qPCR assay was developed for detection of E. ...

Construction of an Affordable and Easy-to-Build Zebrafish Facility

In vivo biomedical research is pivotal to translate in vitro findings into clinical advances. Small academic institutions with limited resources find it virtually impossible to build and maintain typical rodent facilities for research. Zebrafish research has been demonstrated to be a valuable alternative for in vivo research in pharmacology, physiology, development and genetic studies. This article demonstrates that a functional zebrafish facility can be built in an easy and affordable manner. We demonstrate that such a facility could be built in about one working day with minimal tools and expertise. The cost of the 27 1.8 L fish tank zebrafish facility constructed in this study was approximately $1,500. We estimate that the maintenance of an initial working 150 fish colony for 3 months is $1,000. This project involved students, who were introduced to aquaculturing of zebrafish for research proposes.

A Zebrafish facility suitable for breeding and in vivo research is built in an easy and affordable manner. Maintenance is minimal. This facility is ideal for student involvement and independent research.

저렴하고 간편한 구축 Zebrafish의 시설의 건설

In vivo biomedical research is pivotal to translate in vitro findings into clinical advances. Small academic institutions with limited resources find it ...

Plant Promoter Analysis: Identification and Characterization of Root Nodule Specific Promoter in the Common Bean

The upstream sequences of gene coding sequences are termed as promoter sequences. Studying the expression patterns of promoters are very significant in understanding the gene regulation and spatiotemporal expression patterns of target genes. On the other hand, it is also critical to establish promoter evaluation tools and genetic transformation techniques that are fast, efficient, and reproducible. In this study, we investigated the spatiotemporal expression pattern of the rhizobial symbiosis-specific nodule inception (NIN) promoter of Phaseolus vulgaris in the transgenic hairy roots. Using plant genome databases and analysis tools we identified, isolated, and cloned the P. vulgaris NIN promoter in a transcriptional fusion to the chimeric reporter &#946;-glucuronidase (GUS) GUS-enhanced::GFP. Further, this protocol describes a rapid and versatile system of genetic transformation in the P. vulgaris using Agrobacterium rhizogenes induced hairy roots. This system generates &#8805;2 cm hairy roots at 10 to 12 days after transformation. Next, we assessed the spatiotemporal expression of NIN promoter in Rhizobium inoculated hairy roots at periodic intervals of post-inoculation. Our results depicted by GUS activity show that the NIN promoter was active during the process of nodulation. Together, the present protocol demonstrates how to identify, isolate, clone, and characterize a plant promoter in the common bean hairy roots. Moreover, this protocol is easy to use in non-specialized laboratories.

Promoter expression analyses are crucial to improving the understanding of gene regulation and the spatiotemporal expression of target genes. Herein we present a protocol to identify, isolate, and clone a plant promoter. Further, we describe the characterization of the nodule-specific promoter in the common bean hairy roots.

식물 발기인 분석: 식별 및 일반적인 콩에서 루트 결 절 특정 발기인의 특성

The upstream sequences of gene coding sequences are termed as promoter sequences. Studying the expression patterns of promoters are very significant in ...

Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also termed the hypothalamic-pituitary-gonadal axis. The reproductive axis is a key integrator of environmental and internal input adapting fertility to favorable conditions for reproduction. Prior to embarking upon a fertility study in mice and rats, sexual maturity is evaluated to exclude the possibility that the observed reproductive phenotypes are caused by delayed or absent pubertal onset. This protocol describes a non-invasive approach to assess pubertal onset in males through the determination of preputial separation, and in females through vaginal opening and first estrus. After the confirmation of the completion of puberty and the achievement of sexual maturity, a fertility study can be initiated. The procedure describes the optimal breeding conditions for mice and rats, how to set up a fertility study, and what parameters to evaluate and determine if the treatment or gene deletion has an impact on fertility.

Many treatments and genetic mutations impact the timing of sexual maturity and fertility. This protocol describes a non-invasive method to evaluate pubertal onset in mice and rats prior to setting up a fertility study in sexually mature animals.

Pubertal 발병을 확인 하 고 쥐 및 쥐에 다 산 분석 결과 수행 하 여 생식 능력의 결정

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also ...

Loop-Mediated Isothermal Amplification for Screening Salmonella in Animal Food and Confirming Salmonella from Culture Isolation

Loop-mediated isothermal amplification (LAMP) has emerged as a powerful nucleic acid amplification test for the rapid detection of numerous bacterial, fungal, parasitic, and viral agents. Salmonella is a bacterial pathogen of worldwide food safety concern, including food for animals. Presented here is a multi-laboratory-validated Salmonella LAMP protocol that can be used to rapidly screen animal food for the presence of Salmonella contamination and can also be used to confirm presumptive Salmonella isolates recovered from all food categories. The LAMP assay specifically targets the Salmonella invasion gene (invA) and is rapid, sensitive, and highly specific. Template DNAs are prepared from enrichment broths of animal food or pure cultures of presumptive Salmonella isolates. The LAMP reagent mixture is prepared by combining an isothermal master mix, primers, DNA template, and water. The LAMP assay runs at a constant temperature of 65 &#176;C for 30 min. Positive results are monitored via real-time fluorescence and can be detected as early as 5 min. The LAMP assay exhibits high tolerance to inhibitors in animal food or culture medium, serving as a rapid, reliable, robust, cost-effective, and user-friendly method for screening and confirming Salmonella. The LAMP method has recently been incorporated into the U.S. Food and Drug Administration&#8217;s Bacteriological Analytical Manual (BAM) Chapter 5.

Loop-Mediated Isothermal Amplification for Screening Salmonella in Animal Food and Confirming Salmonella from Culture Isolation

Loop-mediated isothermal amplification (LAMP) is an isothermal nucleic acid amplification test (iNAAT) that has attracted broad interest in the pathogen detection field. Here, we present a multi-laboratory-validated Salmonella LAMP protocol as a rapid, reliable, and robust method for screening Salmonella in animal food and confirming presumptive Salmonella from culture isolation.

동물성 식품에서 살모넬라를 선별하고 문화 격리에서 살모넬라를 확인하기 위한 루프 중재 이더말 증폭

Loop-mediated isothermal amplification (LAMP) has emerged as a powerful nucleic acid amplification test for the rapid detection of numerous bacterial, ...