《医学遗传学》课程教学大纲 medical genetics（实验课，英文）

杨北联信大学Medical Genetic Expriment teaching programCourse No. : y700065Course name: Medical Genetics Expriment18credit:1.0Course attribute: requiredCredit hours:Applicablesubject::MedicalGeneticsIntroduction of subjects :Human genetic research involves the study of inherited human traits. Muchof this research is aimed at identifying DNA mutations that can help causespecific health problems, developing methods of identifying thosemutations inpatients,and improving the interventions available to help patients addressthose problems.The identification of genetic mutations enables clinicians topredictthelikelihoodthatpersons will developagivenhealthprobleminthefuture or pass on a health risk to their children. For many disorders, however,there will be a considerable time lag between the ability to determine thelikelihood of disease and the ability to treat the disease.Efforts to isolate DNA mutations involved in disease in order to understandthe origins of the pathophysiological process arebecoming increasingly commonacross the broad sweep of biomedical research, from cardiology to oncology topsychiatry.IRBsshouldexpect to seemore of thesekinds of studies in thefutureVoluntary organizations involved in supporting research on various geneticdisorders.The issues raised in this Section of the Guidebook are addressed withparticular reference to genetic research.Teaching methods: According to the instructional objectives disigningrelevant contents toimprovethemanipulative abilityandbuilduptheinnovationability.knowledge hierarchy: Medical cell biological experiment is the basicexperimental course which focuses on learning several basic technique ofbiological experiment and is the foundation of other courses.、Experimental itemsandarrangementofcourseNO.Contentstypehourrequirment3comfirmationnecessitykaryotype analysis

Medical Genetic Expriment teaching program Course name：Medical Genetics Expriment Course NO.：y700065 Course attribute：required Credit hours： 18 credit：1.0 Applicable subject：：Medical Genetics Introduction of subjects： Human genetic research involves the study of inherited human traits. Much of this research is aimed at identifying DNA mutations that can help cause specific health problems, developing methods of identifying those mutations in patients, and improving the interventions available to help patients address those problems. The identification of genetic mutations enables clinicians to predict the likelihood that persons will develop a given health problem in the future or pass on a health risk to their children. For many disorders, however, there will be a considerable time lag between the ability to determine the likelihood of disease and the ability to treat the disease. Efforts to isolate DNA mutations involved in disease in order to understand the origins of the pathophysiological process are becoming increasingly common across the broad sweep of biomedical research, from cardiology to oncology to psychiatry. IRBs should expect to see more of these kinds of studies in the future. Voluntary organizations involved in supporting research on various genetic disorders. The issues raised in this Section of the Guidebook are addressed with particular reference to genetic research. Teaching methods：According to the instructional objectives disigning relevant contents to improve the manipulative ability and build up the innovation ability. knowledge hierarchy：Medical cell biological experiment is the basic experimental course which focuses on learning several basic technique of biological experiment and is the foundation of other courses. 一、Experimental items and arrangement of course NO. Contents type requirment hour 1 karyotype analysis comfirmation necessity 3

杨北联后大学E6！32comfirmationnecessityBarr body analysis33comfirmationnecessityFingerprint analysisdetection of monogenic43comfirmationnecessitydisease-PCRdetection of monogenic53comfirmationnecessitydisease-Electrophoresis63Human genetic disorder videoscomfirmationnecessityTotal hours16Expriment l: karyotype analysisThe 46 chromosomes located in each somatic cell contain all the genetic materialinherited bythat individual.Locatedinthenucleus,these23pairofhomologouschromosomes are comprisedof 22pair of autosomes (non sex chromosomes) and1pair of sexchromosomes (xX or XY).The genetic material,DNA,exists within the chromosomes andcontains the entire genetic blueprint for the development of an individual.Most normalhuman cells contain identical numbers and types of chromosomes.The analysis of humanchromosomes has allowed researchers to identify the cause of specific genetic diseases andabnormalities.Many genetic disorders have been associated with alterations of the chromosomes anindividual possesses.In someinstances, pieces of chromosomes may be transferred(translocation).On other occasions,pieces of chromosomes maybreak off andbe lostentirely (deletion).Another possibility is that entire chromosomes may be lost or addedAnalyzing an individual's chromosome bytoanindividualschromosomearrangement.doingwhat is called a karyotype can identifyany of these situations.In this exercise, a human tumor cell line,HeLa, isused.TheHeLa cell lineoriginatedin the early 1950' s from the cervical cancer cells of a woman named Henrietta Lacks.Because the cells are of tumor origin, they have continuallydivided and will do so in labcultures for an indefinite period of time.Furthermore, these cells do not contain thenormal diploidnumber of chromosomes of humanbeings(46).Instead theywillpossess achromosome number greater than the diploid number, which is referred to as being aneuploid.Expriment2:BarrbodyanalysisABarrbody(named afterdiscoverer MurrayBarr)istheinactiveX chromosomeinafemalesomatic cell, rendered inactive in a process called lyonization, in those species in whichsex isdeterminedby thepresenceof theY(includinghumans)orWchromosomerather thanthe diploidy of the X or Z. The Lyon hypothesis states that in cells with multiple Xchromosomes, all but one are inactivated during mammalian embryogenesis.This happens early

2 Barr body analysis comfirmation necessity 3 3 Fingerprint analysis comfirmation necessity 3 4 detection of monogenic disease—PCR comfirmation necessity 3 5 detection of monogenic disease—Electrophoresis comfirmation necessity 3 6 Human genetic disorder videos comfirmation necessity 3 Total hours 16 Expriment 1：karyotype analysis The 46 chromosomes located in each somatic cell contain all the genetic material inherited by that individual. Located in the nucleus, these 23 pair of homologous chromosomes are comprised of 22 pair of autosomes (non sex chromosomes) and 1 pair of sex chromosomes (XX or XY). The genetic material, DNA, exists within the chromosomes and contains the entire genetic blueprint for the development of an individual. Most normal human cells contain identical numbers and types of chromosomes. The analysis of human chromosomes has allowed researchers to identify the cause of specific genetic diseases and abnormalities. Many genetic disorders have been associated with alterations of the chromosomes an individual possesses. In some instances, pieces of chromosomes may be transferred (translocation). On other occasions, pieces of chromosomes may break off and be lost entirely (deletion). Another possibility is that entire chromosomes may be lost or added to an individual’s chromosome arrangement. Analyzing an individual’s chromosome by doing what is called a karyotype can identify any of these situations. In this exercise, a human tumor cell line, HeLa, is used. The HeLa cell line originated in the early 1950’s from the cervical cancer cells of a woman named Henrietta Lacks. Because the cells are of tumor origin, they have continually divided and will do so in lab cultures for an indefinite period of time. Furthermore, these cells do not contain the normal diploid number of chromosomes of human beings (46). Instead they will possess a chromosome number greater than the diploid number, which is referred to as being aneuploid. Expriment 2：Barr body analysis A Barr body (named after discoverer Murray Barr) is the inactive X chromosome in a female somatic cell, rendered inactive in a process called lyonization, in those species in which sex is determined by the presence of the Y (including humans) or W chromosome rather than the diploidy of the X or Z. The Lyon hypothesis states that in cells with multiple X chromosomes, all but one are inactivated during mammalian embryogenesis. This happens early

北联后大学inembryonic developmentat random in mammals,except in marsupials and in someextra-embryonictissues of someplacental mammals,in which thefathersX chromosomeisalways deactivated.In men and womenwithmore than one Xchromosome,the numberofBarr bodies visibleat interphase is always one less than the total number of X chromosomes. For example, menwith a 47,xxYkaryotype have a single Barr body,whereas women with a 47,xxxkaryotype havetwo Barr bodies.Barrbodies can be seen on the nucleus of neutrophils.A normal human female has only one Barr body per somatic cell, while a normalhuman male has none.Mammalian X-chromosome inactivation is initiated from the X inactivationcentre or Xic,usually found near the centromere.The center contains twelvegenes, seven of which code for proteins, five for untranslated RNAs, of whichonly two are known to play an active role in the X inactivation process, Xistand Tsix.The centre also appears to be important in chromosome counting:ensuringthatrandominactivation onlytakesplacewhen twoormoreX-chromosomesare present. The provision of an extra artificial Xic in early embryogenesiscan induce inactivation of the single X found in male cells.The roles of Xist and Tsix appear to be antagonistic.Theloss of Tsixexpressionon thefutureinactive Xchromosomeresults inan increase inlevelsof Xist around the Xic. Meanwhile, on the future active X Tsix levels aremaintained:thusthe levels of Xistremain low.Thisshiftallows Xisttobegincoating the future inactive chromosome, spreading out from the Xic. In non-randominactivation this choice appears to be fixed and current evidence suggests thatthe maternally inherited gene may be imprinted.It is thought that this constitutes the mechanism of choice, and allowsdownstream processes to establish the compact state of the Barr body. Thesechanges include histone modifications,such as histone H3 methylation andhistone H2A ubiquitination, [1o] as well as direct modification of the DNA itself,via themethylation of CpG sites.[1i] These changeshelp inactivate geneexpressiononthe inactiveX-chromosomeandtobringaboutitscompactiontoformtheBarrbody.Expriment3:FingerprintanalysisEvery person has minute raised ridges of skin on the inside surfaces of their handsand fingers and on the bottom surfaces of their feet and toes,known as'friction ridgeskin'.The frictionridges provide a gripping surface-in much the same way that the treadpattern of a car tyre does.Friction ridge skin is also the only skin on the bodywithouthairs.Friction ridges do not run evenly and unbroken across our fingers, hands, toes and feet.Rather,they displaya number of characteristicsknown as minutiae.Theprinciple categoriesof minutiae are as follows:A further premise underlying the use of fingerprinting to identify individuals is that

in embryonic development at random in mammals, except in marsupials and in some extra-embryonic tissues of some placental mammals, in which the father's X chromosome is always deactivated. In men and women with more than one X chromosome, the number of Barr bodies visible at interphase is always one less than the total number of X chromosomes. For example, men with a 47,XXY karyotype have a single Barr body, whereas women with a 47,XXX karyotype have two Barr bodies. Barr bodies can be seen on the nucleus of neutrophils. A normal human female has only one Barr body per somatic cell, while a normal human male has none. Mammalian X-chromosome inactivation is initiated from the X inactivation centre or Xic, usually found near the centromere. The center contains twelve genes, seven of which code for proteins, five for untranslated RNAs, of which only two are known to play an active role in the X inactivation process, Xist and Tsix. The centre also appears to be important in chromosome counting: ensuring that random inactivation only takes place when two or more X-chromosomes are present. The provision of an extra artificial Xic in early embryogenesis can induce inactivation of the single X found in male cells. The roles of Xist and Tsix appear to be antagonistic. The loss of Tsix expression on the future inactive X chromosome results in an increase in levels of Xist around the Xic. Meanwhile, on the future active X Tsix levels are maintained; thus the levels of Xist remain low. This shift allows Xist to begin coating the future inactive chromosome, spreading out from the Xic. In non-random inactivation this choice appears to be fixed and current evidence suggests that the maternally inherited gene may be imprinted. It is thought that this constitutes the mechanism of choice, and allows downstream processes to establish the compact state of the Barr body. These changes include histone modifications, such as histone H3 methylation and histone H2A ubiquitination,[10] as well as direct modification of the DNA itself, via the methylation of CpG sites.[11] These changes help inactivate gene expression on the inactive X-chromosome and to bring about its compaction to form the Barr body. Expriment 3：Fingerprint analysis Every person has minute raised ridges of skin on the inside surfaces of their hands and fingers and on the bottom surfaces of their feet and toes, known as 'friction ridge skin'. The friction ridges provide a gripping surface - in much the same way that the tread pattern of a car tyre does. Friction ridge skin is also the only skin on the body without hairs. Friction ridges do not run evenly and unbroken across our fingers, hands, toes and feet. Rather, they display a number of characteristics known as minutiae. The principle categories of minutiae are as follows: A further premise underlying the use of fingerprinting to identify individuals is that

场北联后大学while ridge patterns display immense variability,they can be grouped into patterncategories to facilitate the classification, filing and accessing of very large volumesof fingerprint records. Most English speaking countries have used the'Henry Systemdeveloped by Sir Edward Henry at the end of the nineteenth century. The three basiccategories of ridge patterns in this system are:loops, whorls and arches.Expriment 4-5:detection of monogenic diseasePCR and ElectrophoresisHuntington's disease is caused by a genetic defect on chromosome #4. The defect causesa part of DNA, called a CAG repeat, to occur many more times than it is supposed to. Normally,this section of DNA is repeated 10 to 35 times. But in persons with Huntington's disease,itisrepeated36 to 120times.As thegene ispassed onfromone generationto thenext,the number of repeats - called a CAG repeat expansion -tend to get larger. The larger thenumber of repeats, the greater your chance of developing symptoms at an earlier age.Developedin1984byKaryMullis,PCRisnowacommonandoftenindispensabletechniqueused in medical and biological research labs for a variety of applications.These includeDNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; thediagnosis of hereditary diseases; the identification of genetic fingerprints (used inforensic sciences and paternity testing);and the detection and diagnosis of infectiousdiseases.In 1993 Mullis was awarded the Nobel Prize in Chemistry for his work on PCR.Agarose gel electrophoresis can resolve molecules based on charge, size, and shape.Agarose Gel Electrophoresisisusedto separate,identify,andpurifyDNAfragments.Thistechnique is simple, rapid to perform. Furthermore, agarose gel can be poured in a varietyof shapes, sizes, and porosities and can be run in a number of different configurations.If necessary, these bands of DNA can be recovered from the gel and used for a variety ofpurposes.Expriment 6:Human genetic disorder videosFor most of genetic disorder patients can' t easily to see ,we can to knowhow the genetic disorders was from some videos.examinationmode:closed-book examandday-to-dayclasswork

while ridge patterns display immense variability, they can be grouped into pattern categories to facilitate the classification, filing and accessing of very large volumes of fingerprint records. Most English speaking countries have used the 'Henry System' developed by Sir Edward Henry at the end of the nineteenth century. The three basic categories of ridge patterns in this system are: loops, whorls and arches. Expriment 4-5：detection of monogenic disease—PCR and Electrophoresis Huntington's disease is caused by a genetic defect on chromosome #4. The defect causes a part of DNA, called a CAG repeat, to occur many more times than it is supposed to. Normally, this section of DNA is repeated 10 to 35 times. But in persons with Huntington's disease, it is repeated 36 to 120 times.As the gene is passed on from one generation to the next, the number of repeats - called a CAG repeat expansion - tend to get larger. The larger the number of repeats, the greater your chance of developing symptoms at an earlier age. Developed in 1984 by Kary Mullis, PCR is now a common and often indispensable technique used in medical and biological research labs for a variety of applications. These include DNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic fingerprints (used in forensic sciences and paternity testing); and the detection and diagnosis of infectious diseases. In 1993 Mullis was awarded the Nobel Prize in Chemistry for his work on PCR. Agarose gel electrophoresis can resolve molecules based on charge, size, and shape. Agarose Gel Electrophoresis is used to separate, identify, and purify DNA fragments. This technique is simple, rapid to perform. Furthermore, agarose gel can be poured in a variety of shapes, sizes, and porosities and can be run in a number of different configurations. If necessary, these bands of DNA can be recovered from the gel and used for a variety of purposes. Expriment 6：Human genetic disorder videos For most of genetic disorder patients can’t easily to see ,we can to know how the genetic disorders was from some videos. examination mode： closed-book exam and day-to-day class work