A M TO-34 C- C+ TO-34 C C+ B Probe (0.96 kb) Figure 4. Southern blot hybridisation analysis of genomic DNA extracted from leaves of two transgenic cotton lines and the non-transformed plant (BRS 293). (A) DNA digested with Hind/ll; (B) blot exposition in X-ray. C- - negative control: DNA from BRS 293 digested with Hind III; C+ - positive control: DNA from BRS 293 digested with Hind III+ probe (0.93 kb probe, 1 ng µL−¹); M – 1 kb molecular marker (Ladder Plus; Invit- rogen).

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
icon
Related questions
icon
Concept explainers
Question
100%
Use the information provided in the experiment below to annotate/ label the figure. Background: Boll weevil is a serious pest of cotton crop. Effective control involves applications of chemical insecticides, increasing the cost of production and environmental pollution. The current genetically modified Bt crops have allowed great benefits to farmers but show activity limited to lepidopteran pests. This work reports on procedures adopted for integration and expression of a cry transgene conferring resistance to boll weevil and fall armyworm by using molecular tools. Four Brazilian cotton cultivars were microinjected with a minimal linear cassette generating 1248 putative lines. Complete gene integration was found in only one line (TO-34) containing one copy of crylla detected by Southern blot. Protein was expressed in high concentration at 45 days after emergence (dae), decreasing by approximately 50% at 90 dae. Toxicity of the cry protein was demonstrated in feeding bioassays revealing 56.7% mortality to boll weevil fed buds and 88.1% mortality to fall armyworm fed leaves. A binding of crylla antibody was found in the midgut of boll weevils fed on TO-34 buds in an immunodetection assay. The gene introduced into plants confers resistance to boll weevil and fall armyworm. Transmission of the transgene occurred normally to TI progeny. All plants showed phenotypically normal growth, with fertile flowers and abundant seeds. Results relating to figure: A total of 1248 putative transgenic seeds were collected from microinjected bolls of four Brazilian cultivars and sown in the greenhouse. At first, PTLs were analysed by PCR using three primer combinations. A few amplicons were verified in the cultivars and, even so, in one or two primer combinations, indicating incomplete integration of the gene. Only in cv. BRS 293 was at least one PTL (TO-34) identified in all primer combinations, indicating possible complete integration of crylla based on PCR reactions, mainly with the 5 F/2R primer combination, which generated an amplicon corresponding to the complete gene sequence (2.1 kb). A fragment of 0.44 kb (1 F/3R) collected from this line was purified, sequenced and analysed using BLAST tools.The results showed high between the sequences of crylla from BRS 293 T0-34 and the Bt cry1Ia gene deposited in the NCBI gene bank (DQ535488, E-value: 3e−63 to 1e−12). In order to confirm the number of copies of cry1Ia integrated to the genome of the T0-34 line, Southern blot assays were carried out using a DIG-labelled probe from the cry1Ia gene (0.96 kb). Only one hybridisation signal was seen at∼5.0 kb (Fig. 4), indicating that complete gene construction was successfully introduced.
A
M TO-34 C- C+
TO-34 C C+
B
Probe (0.96 kb)
Figure 4. Southern blot hybridisation analysis of genomic DNA extracted
from leaves of two transgenic cotton lines and the non-transformed
plant (BRS 293). (A) DNA digested with HindIll; (B) blot exposition in
X-ray. C- - negative control: DNA from BRS 293 digested with Hind III;
C+ - positive control: DNA from BRS 293 digested with Hind III + probe
(0.93 kb probe, 1 ng µL-¹); M - 1 kb molecular marker (Ladder Plus; Invit-
rogen).
Transcribed Image Text:A M TO-34 C- C+ TO-34 C C+ B Probe (0.96 kb) Figure 4. Southern blot hybridisation analysis of genomic DNA extracted from leaves of two transgenic cotton lines and the non-transformed plant (BRS 293). (A) DNA digested with HindIll; (B) blot exposition in X-ray. C- - negative control: DNA from BRS 293 digested with Hind III; C+ - positive control: DNA from BRS 293 digested with Hind III + probe (0.93 kb probe, 1 ng µL-¹); M - 1 kb molecular marker (Ladder Plus; Invit- rogen).
Expert Solution
trending now

Trending now

This is a popular solution!

steps

Step by step

Solved in 3 steps

Blurred answer
Knowledge Booster
Molecular techniques
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.
Similar questions
  • SEE MORE QUESTIONS
Recommended textbooks for you
Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
Biology
ISBN:
9780134580999
Author:
Elaine N. Marieb, Katja N. Hoehn
Publisher:
PEARSON
Biology 2e
Biology 2e
Biology
ISBN:
9781947172517
Author:
Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:
OpenStax
Anatomy & Physiology
Anatomy & Physiology
Biology
ISBN:
9781259398629
Author:
McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa Stouter
Publisher:
Mcgraw Hill Education,
Molecular Biology of the Cell (Sixth Edition)
Molecular Biology of the Cell (Sixth Edition)
Biology
ISBN:
9780815344322
Author:
Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter
Publisher:
W. W. Norton & Company
Laboratory Manual For Human Anatomy & Physiology
Laboratory Manual For Human Anatomy & Physiology
Biology
ISBN:
9781260159363
Author:
Martin, Terry R., Prentice-craver, Cynthia
Publisher:
McGraw-Hill Publishing Co.
Inquiry Into Life (16th Edition)
Inquiry Into Life (16th Edition)
Biology
ISBN:
9781260231700
Author:
Sylvia S. Mader, Michael Windelspecht
Publisher:
McGraw Hill Education