New User Special Price Expires in

Let's log you in.

Sign in with Facebook


Don't have a StudySoup account? Create one here!


Create a StudySoup account

Be part of our community, it's free to join!

Sign up with Facebook


Create your account
By creating an account you agree to StudySoup's terms and conditions and privacy policy

Already have a StudySoup account? Login here


by: Renee Lehner


Renee Lehner
GPA 3.73


Almost Ready


These notes were just uploaded, and will be ready to view shortly.

Purchase these notes here, or revisit this page.

Either way, we'll remind you when they're ready :)

Preview These Notes for FREE

Get a free preview of these Notes, just enter your email below.

Unlock Preview
Unlock Preview

Preview these materials now for free

Why put in your email? Get access to more of this material and other relevant free materials for your school

View Preview

About this Document

Class Notes
25 ?




Popular in Course

Popular in Biology

This 19 page Class Notes was uploaded by Renee Lehner on Wednesday September 9, 2015. The Class Notes belongs to BIOL 411 at University of Washington taught by Staff in Fall. Since its upload, it has received 10 views. For similar materials see /class/192341/biol-411-university-of-washington in Biology at University of Washington.


Reviews for DEV BIOL


Report this Material


What is Karma?


Karma is the currency of StudySoup.

You can buy or earn more Karma at anytime and redeem it for class notes, study guides, flashcards, and more!

Date Created: 09/09/15
Chapter 4 The genetic core of development I What do genes have to do with how an organism progresses from fertilized egg to adult A Genomic equivalence B differential gene expression Genomic equivalence that every cell has the same nuclear genes so how do cells perform speci c functions and acquire diff fates Use cloning techniques to determine ifwhen nuclear potency is restricted transferred nucleus into egg from a donor cell eg a differentiated muscle cell can compare results from using donor cells at diff dev stages 11 How do we then understand the control of differential gene expression must use molecular techniques to uncover structure and function of DNA RNA and proteins 1 RNA localization Northern blotting 1 39 I RTPCR microarrays insitu hybridization 999 a Northern blotting can determine when dev stage and where tissue driferem s are expressed Protocol Extract RNA NA electorphoresi Transfer RNAs to nitrocellulose Separate by size w gel a 5 t Fquot 1 Results if mRNA for gene is in sample the labeled DNA will bind and be detected by autoradiography b Reverse transcriptase polymerase chain reaction RTPCR First quick review of PCR amplification wzimed gene rgt35ihuch immnm 2 6x hiilmn mm mm mm m Reverse transcriptase polymerase chain reaction RTPC R can be dif cult to extract enough RNA for molecular assays get around this by making cDNA from mRNA l 5 mRNA 3 l Anneal primal l Reverse transcripmse mRVA u my CD A l Mkali m w eDNA c Microarrays to determine changes in transcription of many genes In developmental biology DNA microarrays have already been used to 1 compare gene expression for the entire Drosophila larva during metamorphosis White KP Rifkin SA Hurban P Hogness DS 1999 Microarray analysis of Drosophila development during metamorphosis Science 286 217984 2 look at which genes are upregulated or downregulated when embryonic stem cells are caused to differentiate in culture Kelly DL and Rizzino A 2000 DNA microarray analyses of genes regulated during the differentiation of embryonic stem cells Mol Reprod Dev 56 113123 3 compare the gene expression in activated vs unactivated B and T cells Ollila J and Vihinen M 1998 Stimulation of B and T cells activates expression of transcription and differentiation factors Biochem Biophys Res Commun 249 475 480 Procedure for microarray analysis 1 Isolate mRNA from organism at one stage eg gastrula or from one cell type use reverse transcriptase to generate cDNA and label with green fluorescence 2 Isolate mRNA from organism at another stage eg neurula or second cell type use RT to generate cDNA and label with red fluorescence 3 Add hybridize both cDNA probes to chip has all cDNA of genome and wash Prepare Mgctoarray a b m J A N D c m a D m E d insitu hybridization generate antisense mRNA from a target cloned gene where the gene is reversed so the product transcribed is complementary to the gene s normal product antisense mRNA is labeled cells labeled contain the mRNAs of the target gene Ctrl hm p7quot mxh wm M Us bupr an RNA burpBb mRNA B I C D E liturqu 1 icw M l yng 4 Mgr f J tl jh sll mic nil Hans S et al BMC Dev Biol 2007 d insitu hybridization A cDNA compleaemary Alkaline phusplmtasu Colorless compound to speci c mRNA cnnjugatcd antibudy that becomes purple MT Hilles in the cell 0 dig gmi dyr when phosphalc made by detergent 15 removcd Cell 7 x membrane a Dlgoxirgemn label on uddine Wash P IIIHN I mam 1 Add digoxigj39iinlabcled pmbc 2 Add alkaline phplmam conjugated 3 Add chemical that becomes a dark complementary to RNA of internsl amibody 11m binds to digoxigenin purple dye when phmphate is removed dye colors Lhc cell 11 How do we then understand the control of differential gene expression must use molecular techniques to uncover structure and function of DNA RNA and proteins 2 Determining function of genes a Transgenic cellsorganisms b Antisense RNA a Transgenic cellsorganisms 1 Introducing genes into cells to study their function use microinjection to introduce cloned gene into cell can also use transfection to incorporate DNA into cells the cells take up the DNA or use electroporation to force DNA into cells w electrical current 39 39 Roszko I et al Dev Biol 2006 a Transgenic cellsorganisms 1 Introducing genes into cells to study their function W W transposable elements mobile 5 Tmnspositionleamg DNA that Jump around In Eig39gijtied frsstnmend genome interupt genes and quot7 can use to study normal gene function and can use for inserting genes in ectopic Homologoussequeme regions of genome G azgfip iratemp39a m Examples 1 Pelements in Drosophila m 2 TD NA in Arabidopsis Completion of repair C replaces original P element with newly synthesized copy H E a Transgenic cellsorganisms 2 Chimeric animals take advantage of embryonic stem cells and totipotency and techniques to incorporate DNA into cells from previous slide use to make transgenic animals with GFP fused to gene of interest to follow expression through development a Transgenic cellsorganisms 3 Gene knockout Mae lumr disrupt at transcriptional gmhrgm level Tultur ul 3 I15 Eleclmpanllun m 8 Em En gt Rtsu39iction Lapse Mum1d 8MP rmdrmudm um m 5cm inscniun en hmmzygo 5 ES ILA m Ihcir nrmnydn nsisimc Iujcn hulrmlygoui Ls cell inm blmmqn lnim blasnxym mm mm anmimi a chimeric mite All Pg Oi xd Q Q Brcdcllimurinuwildmk Mm lmLmzylrous QBMPTIBMPT vld Mummy 2 Htlrmxygalc KMW JBMPT BMPT IHMPT39 HMPF39IBMIW X Wild type llnmurmu Hclcmvrgmc HI 7 IBMP7 EMPTIFMPT BMW IHMW39 Knockout phenotype examples H II n39ilxlrhpr b Functions of the gene message 1 Antisense RNA Create by putting cloned DNA into vectors that have promoters at either end of gene and incubate vector with RNA pol eg T3T7 pol to read in reverse direction antisense sequence Inject antisense mRNA into animal and this will bind to endogenous RNA and inhibit its function A in Kriippul mutmt embryo same Q v m i I 7 P 7 promoter VViidiypc embryo QIEHM Wildmyc embryo injected with Krrlppel amiscnse RNA T3 39 yquot T7 pmmum zglt polymerase V actor DNA 4 b Functions of the gene message 2 RNA interference use dsRNA to degrade the targeted message does NOT a inhibit translation mill rim vnn FTP 151 1 can inject dsRNA into animal 2 These dsRNAs activate the enzyme Dicer that cuts the RNA into small pieces 3 The siRNAs bind to the RISC complex that unwinds the strands so that the siRNAs can bind the complementary endogenous RNAs which are degraded Motegi and Sugimoto Nat Cell Biol 2006


Buy Material

Are you sure you want to buy this material for

25 Karma

Buy Material

BOOM! Enjoy Your Free Notes!

We've added these Notes to your profile, click here to view them now.


You're already Subscribed!

Looks like you've already subscribed to StudySoup, you won't need to purchase another subscription to get this material. To access this material simply click 'View Full Document'

Why people love StudySoup

Steve Martinelli UC Los Angeles

"There's no way I would have passed my Organic Chemistry class this semester without the notes and study guides I got from StudySoup."

Janice Dongeun University of Washington

"I used the money I made selling my notes & study guides to pay for spring break in Olympia, Washington...which was Sweet!"

Jim McGreen Ohio University

"Knowing I can count on the Elite Notetaker in my class allows me to focus on what the professor is saying instead of just scribbling notes the whole time and falling behind."

Parker Thompson 500 Startups

"It's a great way for students to improve their educational experience and it seemed like a product that everybody wants, so all the people participating are winning."

Become an Elite Notetaker and start selling your notes online!

Refund Policy


All subscriptions to StudySoup are paid in full at the time of subscribing. To change your credit card information or to cancel your subscription, go to "Edit Settings". All credit card information will be available there. If you should decide to cancel your subscription, it will continue to be valid until the next payment period, as all payments for the current period were made in advance. For special circumstances, please email


StudySoup has more than 1 million course-specific study resources to help students study smarter. If you’re having trouble finding what you’re looking for, our customer support team can help you find what you need! Feel free to contact them here:

Recurring Subscriptions: If you have canceled your recurring subscription on the day of renewal and have not downloaded any documents, you may request a refund by submitting an email to

Satisfaction Guarantee: If you’re not satisfied with your subscription, you can contact us for further help. Contact must be made within 3 business days of your subscription purchase and your refund request will be subject for review.

Please Note: Refunds can never be provided more than 30 days after the initial purchase date regardless of your activity on the site.