Rekombinasi DNA

Dosen : Faizah Fulyani, S.Si, M.Sc, Ph.D

2018

Referensi :
1. Biologi Molekuler, Triwibowo Yuwono
2. Essential Cell Biology, Bruce Albert
3. Video pembelajaran
4. PERHATIAN: slide ini merupakan alat bantu dalam perkuliahan dimana penjelasan
detail anda dapatkan dalam proses perkuliahan. Bacalah textbook yang disarankan di
atas dan catatan yang anda dibuat selama proses perkuliahan.

Disclaimer: All Materials in this presentation are belong to their respective owner and used for educational purposes only.
 Biological Roles for Recombination

1. Generating new gene/allele combinations

(crossing over during meiosis)
2. Generating new genes (e.g., Immuno-
globulin rearrangement)
3. Integration of a specific DNA element (or
virus)
4. DNA repair

Practical use: Making transgenic cells and

organisms
 Types of Recombination

1. Homologous - occurs between sequences

that are nearly identical (e.g., during
meiosis)
2. Site-Specific - occurs between sequences
with a limited stretch of similarity; involves
specific sites
3. Transposition – DNA element moves from
one site to another, usually little sequence
similarity involved
 Homologues recombination
• often occurs shortly after a cell’s DNA has
been replicated before cell division, when the
duplicated helices are still physically close to
each other
• 2 ds DNA that have similar not necessarily
identical region are align then cross over.
• The site exchange can occur anywhere
• No nucleotide sequence are altered at site
exchange (no lost nor gained)
 Homologues recombination

Basic common steps of homologues recombination

DSB

3’-overhang
formation
Strand
invasion

Sumber gambar: skolvo & MSU

Homologues recombination
Basic common steps of homologues recombination

Extension/
strand
displacement

Invasion of
the second
strand
Holliday
junction
Homologues recombination

Basic common steps of homologues recombination

Strand
Symmetry of the Holliday junction invasion
Homologues recombination
Basic common steps of homologues recombination
RuvC

RuvC

gene conversion crossingover

 Homologues recombination
Source of DSB
Radiation/chemical DSB

Single strand break in the template strand during replication

1. Uraikan tahapan-tahapan dalam proses
rekombinasi homolog DNA (10 ‘) yang terdapat
dalam video berikut ini.

Formation of Holliday junction

 2. Uraikan tahapan dan enzim yang terlibat
dalam proses rekombinasi pada bakteri (10 ‘)

Homologues recombination
Recombination in bacteria
 Meiosis
Meiotic recombination aims at genetic diversity

2N (diploid) 2N (diploid)
chromosome set chromosome set
somatic cells maturation of
sperm and egg 1N (haploid)
chromosome set
sperm and egg
 Meiosis
Meiotic recombination aims at genetic diversity

fertilization
replication

2N (diploid)
chromosome set
1N (haploid) somatic cells
chromosome set
sperm and egg
 Meiosis
Profase 1 of Meiosis

Meiosis I

leptotene zygotene pachytene diplotene diakinesis

chromosome chromosome crossing chromosomes separation of
condensation pairing over bound by chromosomes
chiasmata
Meiosis II
 Meiosis
Regulation of crossingover

Crossingover events
are tightly regulated

No crossingover between sister

chromatids
(S. cerevisiae)

One crossingover per chromosome

(strictly - C. elegans,
relaxed - other eukarya)
VDJ recombination
Structure of antibody

Variable Antigen
regions binding

light
chain

Constant
regions
heavy
chain
 VDJ recombination
Light chain diversity generation
Vκ Jκ
V1 V2 V3 V76 J1-J5 Cκ

>150 variants recombination

V1 V2 V3J4-J5 Cκ

transcription

V3J4-J5 Cκ

splicing

V3J4 Cκ
 VDJ recombination
Heavy chain diversity generation
VH DΗ JΗ CΗ
V1 V2 V3 V50 D1-D30 J1-J6 Cµ ..... Cα

>9000 variants recombination

V1 V2D15 J3-J6 Cµ ..... Cα

transcription

V2D15 J3-J6 Cµ ..... Cα

150х9000=10
6
splicing
functional variants
V2D15J3 Cµ
DNA recombination
technology
Source: nlm.nih.gov
Selamat Belajar