kuliah kloning
DESCRIPTION
imunologiTRANSCRIPT
![Page 1: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/1.jpg)
Kloning Gen
![Page 2: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/2.jpg)
Pendahuluan: BioteknologiBiotechnology
Dolly and surrogate Mom
Embryonic stem cells and gene therapy Genetically modified rice.
![Page 3: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/3.jpg)
BiotechnologyBioteknologi secara luas didefinisikan sebagai rekayasa organisme untuk mendapatkan kemanfaatan tertentu.
Bioteknologi seringkali melibatkan modifikasi dan hibridisasi gen dan pemasukan pada organisme tertentu, dimana secara normal gen tersebut tidak ditemukan dalam organisme tersebut.
Fourteen month-old genetically engineered (“biotech”) salmon (left) and standard salmon (right).
![Page 4: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/4.jpg)
Aplikasi Biotechnology
Kloning binatang
Kloning gen untuk produksi hormon
Pengembangan stem sel DNA fingerprinting
Perdebatan etik?
![Page 5: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/5.jpg)
Kloning untuk reproduksi
Even under the best of circumstances, the current technology of cloning is very inefficient.
Cloning provides the most direct demonstration that all cells of an individual share a common genetic blueprint.
![Page 6: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/6.jpg)
The Stem Cell Concept
A stem cell is an undifferentiated, dividing cell that gives rise to a daughter cell like itself and a daughter cell that becomes a specialized cell type.
![Page 7: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/7.jpg)
Stem Cells are Found in the Adult, but the Most Promising Types of Stem Cells for Therapy are Embryonic Stem Cells
![Page 8: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/8.jpg)
The Inner Cell Mass is the Source of Embryonic Stem Cells
The embryo is destroyed by separating it into individual cells for the collection of ICM cells.
![Page 9: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/9.jpg)
In order to obtain self-derived stem cells, adult cell nuclei are transferred to human oocytes, obtained as excess material from in-vitro fertilization.This procedure was used to create the sheep “Dolly”, and is at present highly controversial, and ethically debated, as the resulting blastula from the procedure could be transferred to a human foster mother, and give rise to a complete human being. Figure reproduced by permission from Solter et al. ScienceMag.
Human therapeutic cloning.
![Page 10: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/10.jpg)
Kloning - definisi Dari Bahasa Yunani - klon, ranting Kumpulan turunan suatu individu yang
dihasilkan tanpa melalui perkawinan; kumpulan replika sebagian atau seluruh makromolekul (contoh, DNA atau antibodi)
Suatu individu yang tumbuh dari satu sel somatik induknya serta memiliki identitas genetik yang sama dengan induknya
Klon: Koleksi molekul atau sel yang semua identitasnya sama dengan molekul atau sel penurunnya
![Page 11: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/11.jpg)
Kloning DNA
Metoda untuk memurnikan atau mengidentifikasi dan memperbanyak suatu potongan DNA tertentu (klon) yang dikehendaki dari campuran potongan-potongan DNA yang kompleks.
![Page 12: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/12.jpg)
Kloning Gen
Ketika keseluruhan DNA dari suatu organisme diekstraksi, akan diperoleh seluruh gen yang dimiliki organisme tersebut
Pada kloning gen, hanya gen (DNA) tertentu yang diisolasi, dimurnikan, dan diperbanyak (diklon)
![Page 13: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/13.jpg)
Tujuan mengklon Gen Menentukan urutan basa nukleotida penyusun gen tersebut
Menganalisis atau mengidentifikasi urutan basa nukleotida pengendali gen tersebut
Mempelajari fungsi RNA / protein/enzim yang disandi gen tersebut
Mengidentifikasi mutasi yang terjadi pada kecacatan gen yang mengakibatkan penyakit bawaan
Merekayasa organisme untuk tujuan tertentu, misalnya memproduksi insulin, ketahanan terhadap hama, dll.
![Page 14: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/14.jpg)
Sumber DNA untuk diklon DNA kromosom cDNA (complementary DNA) yang
disintesis menggunakan mRNA sebagai cetakan (template)
DNA yang dihasilkan dari perbanyakan menggunakan PCR
![Page 15: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/15.jpg)
Mensintesis cDNA
![Page 16: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/16.jpg)
Perbanyakan DNA dengan PCR (Polymerase Chain Reaction)
![Page 17: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/17.jpg)
Bahan / Alat untuk Mengklon
Enzim endonuklease restriksi Enzim ligase Vektors Inang (Host) Metoda untuk memasukkan DNA ke dalam
sel inang
![Page 18: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/18.jpg)
Memotong DNA Menggunakan enzim
endonuklease restriksi Ujung “lengket” (sticky
ends) Ujung “tumpul” (blunt
ends) Penamaan enzim
EcoRI E = genus (Escherichia) co = species (coli) R = strain I = # of enzyme
![Page 19: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/19.jpg)
Ujung “lengket” dan “tumpul” (Blunt & Sticky ends)
![Page 20: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/20.jpg)
Penyambungan (pasting) DNA
Pembentukan ikatan-H pada ujung-ujung yang komplemen (sticky ends)
Ligase membentuk ikatan fosfodiester untuk merekatkan benang-benang DNA
![Page 21: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/21.jpg)
![Page 22: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/22.jpg)
Vektor untuk MengklonDiperlukan suatu wahana (vehicle) untuk memasukkan suatu potongan DNA ke dalam sel agar DNA tersebut dapat disimpan dan diperbanyak di dalam sel tersebut
![Page 23: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/23.jpg)
PlasmidDNA bukan kromosom (extrachromosomal
DNA) yang secara alami dimiliki suatu jasad
Bentuknya benang ganda (double strands DNA, dsDNA) sirkular
Plasmid buatan (Artificial plasmids) dapat dibuat dengan menambahkan potongan-potongan DNA lain
![Page 24: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/24.jpg)
Plasmid dapat dimodifikasi untuk mampu membawa potongan DNA lain ke dalam sel bila memiliki:
Replikator (origin of replication) Penanda (Marker) yang mudah diseleksi
(misalnya gen ketahanan terhadap antibiotik)
Situs untuk mengklon (potongan DNA yang memiliki urutan basa nukleotida yang menjadi sasaran enzim restriksi tetapi tidak terletak di dalam daerah replikator atau penanda
Vektor untuk Mengklon
![Page 25: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/25.jpg)
Plasmid yang Dimiliki oleh Escherichia coli
Berasal dari plasmid alami E. coli
Potongan DNA tambahan
Potongan DNA tambahan
![Page 26: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/26.jpg)
![Page 27: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/27.jpg)
Plasmid Khimera (Chimeric Plasmids)
Khimera berasal dari mitologi Yunani, makhluk dengan tubuh gabungan dari bagian-bagian makhluk binatang lain
Setelah pemotongan plasmid menggunakan suatu enzim restriksi, potongan DNA asing yang memiliki ujung pemotongan yang sama dapat disisipkan
Setelah ujung-ujung plasmid dan potongan DNA asing disambung, akan dihasilkan "plasmid rekombinan"
Plasmid rekombinan dapat bereplikasi dalam sel inang yang sesuai
![Page 28: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/28.jpg)
![Page 29: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/29.jpg)
Kloning TerorientasiBila diinginkan untuk
menginsersikan potongan DNA asing dengan orientasi tertentu
Dilakukan dengan memotong DNA vektor maupun DNA sumber gen yang dikehendaki menggunakan dua enzim restriksi yang berbeda
![Page 30: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/30.jpg)
![Page 31: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/31.jpg)
Vektor untuk Mengklon1 Vektor berupa plasmid 2 Vektor berupa bakteriofaga3 Cosmid 4 BACs (Bacterial Artificial
Chromosome) & YAC (Yeast Artificial Chromosome)
![Page 32: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/32.jpg)
1. Memiliki origin of replication dari inang yang dituju, sehingga memungkinkan replikasi secara independen terhadap genom inang.
2. Memiliki penanda selektif: Memudahkan seleksi sel pembawa plasmid tersisipi DNA asingketahanan terhadap antibiotik ganda penapisan biru-putih
3. Memiliki banyak situs pengkloningan (multiple cloning sites, MCS)
4. Mudah diisolasi dari sel inang.
Vektor berupa Plasmid
![Page 33: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/33.jpg)
Multiple Cloning Site (MCS)
![Page 34: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/34.jpg)
Vektor berupa Plasmid
![Page 35: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/35.jpg)
Vektor berupa Plasmid Keunggulan: Kecil, mudah pengerjaannya Strategi seleksi mudah Berguna untuk mengklon potongan
DNA ukuran kecil (< 10kbp) Kelemahan:
Kurang bermanfaat untuk mengklon potongan DNA ukuran besar (> 10kbp)
![Page 36: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/36.jpg)
Memilih Vektor Ukuran DNA yang
disisipkan Ukuran vektor Situs enzim
restriksi yang tersedia
Jumlah salinan (copy number)
Efisiensi kloning Kemampuan untuk
menapis DNA sisipan
Rencana penelitian selanjutnya
![Page 37: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/37.jpg)
Cara Mengklon DNA (1) Isolasi vektor kloning
(plasmid bacterial) & DNA sumber gen
Pemotongan DNA sumber gen & vektor kloning menggunakan enzim restriksi yang sama
Penyisipan potongan DNA sumber gen ke dalam vektor kloning yang telah dipotong menggunakan enzim restriksi yang sama; potongan disambung dengan bantuan enzim DNA ligase
![Page 38: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/38.jpg)
Cara Mengklon DNA (2) Vektor kloning yang
telah tersisipi potongan DNA dimasukkan ke dalam sel inang (transformasi sel inang)
Penapisan sel pengklon (dan gen yang dimasukkan)
Identifikasi sel pengklon pembawa gen yang dikehendaki
![Page 39: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/39.jpg)
Memudahkan seleksi sel pembawa plasmid tersisipi DNA asing
ketahanan terhadap antibiotik ganda
penapisan biru-putih
Penanda selektif
![Page 40: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/40.jpg)
Penapisan Klon Medium pertumbuhan
diberi antibiotik yang sesuai dengan sifat ketahanan yang digunakan sebagai penanda, misalnya Kanamisin
Bakteri di paruh cawan petri sebelah kanan memiliki plasmid dengan penanda ketahanan terhadap Kanamisin(Kanr), yang di sebelah kiri tidak memilikinya
![Page 41: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/41.jpg)
Penapisan warna koloni Biru/Putih
lacZ insert
Enzim tidak berfungsi
X-gal produk
lacZ
Enzim berfungsi
X-gal produk
![Page 42: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/42.jpg)
Route to the Production by Bacteria of Human Insulin
![Page 43: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/43.jpg)
![Page 44: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/44.jpg)
Apakah problemnya?
DNA mamalia mengalami splicing pada proses transkripsi?
Bagaimana solusinya? Gunakan cDNA
![Page 45: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/45.jpg)
![Page 46: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/46.jpg)
Penelusuran melalui BLAST
We can enter the primary structure of insulin protein into a database (“BLAST” : [Basic Local Alignment Search Tool]) which will compare the protein sequence with the DNA sequence of the human genome. It will identify the DNA that is capable of coding the insulin protein and tell us which DNA molecule this is.
![Page 47: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/47.jpg)
Primary structure of insulin protein :NMALWMRLLPLLALLALWGPDPAAAFVNQHL
CGSHLVEALYLVCGERGFFYTPKTRREAEDLQVGQVELGGGPGAGSLQPLALEGSLQKRGIVEQCCTSICSLYQLENYCN -C
![Page 48: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/48.jpg)
Urutan m-RNANM_000207 Reports Homo sapiens insu...[gi:109148525]Links>gi|109148525|ref|NM_000207.2| Homo sapiens insulin (INS), mRNA5’AGCCCTCCAGGACAGGCTGCATCAGAAGAGGCCATCAAGCAGATCACTGTCCTTCTGCCATGGCCCTGTGGATGCGCCTCCTGCCCCTGCTGGCGCTGCTGGCCCTCTGGGGACCTGACCCAGCCGCAGCCTTTGTGAACCAACACCTGTGCGGCTCACACCTGGTGGAAGCTCTCTACCTAGTGTGCGGGGAACGAGGCTTCTTCTACACACCCAAGACCCGCCGGGAGGCAGAGGACCTGCAGGTGGGGCAGGTGGAGCTGGGCGGGGGCCCTGGTGCAGGCAGCCTGCAGCCCTTGGCCCTGGAGGGGTCCCTGCAGAAGCGTGGCATTGTGGAACAATGCTGTACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAACTAGACGCAGCCCGCAGGCAGCCCCACACCCGCCGCCTCCTGCACCGAGAGAGATGGAATAAAGCCCTTGAACCAGCAAAA3’*although this is a mRNA sequence, it is shown as having “T”s andnot “U”s. This is because the sequence information was generatedfrom a DNA molecule that was made in the laboratory by using theenzyme “Reverse Transcriptase”. This DNA was “complementary” tothe mRNA, and is called “complementary DNA” or “cDNA”.
![Page 49: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/49.jpg)
NM_000207. Reports Homo sapiens insu...[gi:109148525]Links>gi|109148525|ref|NM_000207.2| Homo sapiens insulin (INS),
mRNA5’AGCCCTCCAGGACAGGCTGCATCAGAAGAGGCCATCAAGCAGA
TCACTGTCCTTCTGCCATGGCCCTGTGGATGCGCCTCCTGCCCCTGCTGGCGCTGCTGGCCCTCTGGGGACCTGACCCAGCCGCAGCCTTTGTGAACCAACACCTGTGCGGCTCACACCTGGTGGAAGCTCTCTACCTAGTGTGCGGGGAACGAGGCTTCTTCTACACACCCAAGACCCGCCGGGAGGCAGAGGACCTGCAGGTGGGGCAGGTGGAGCTGGGCGGGGGCCCTGGTGCAGGCAGCCTGCAGCCCTTGGCCCTGGAGGGGTCCCTGCAGAAGCGTGGCATTGTGGAACAATGCTGTACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAACTAGACGCAGCCCGCAGGCAGCCCCACACCCGCCGCCTCCTGCACCGAGAGAGATGGAATAAAGCCCTTGAACCAGCAAAA3’
5’-ATGGCCCTGT….. TACTGCAACTAG-3’ :is the ORF (protein coding region) of insulin cDNA and is theregion we need to make insulin protein in the laboratory
![Page 50: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/50.jpg)
Amplifikasi cDNA
Before we can clone the insulin gene into the plasmid, wehave to make large quantities of the insulin gene (thesequence corresponding to the ORF of the insulin mRNA =cDNA) in the laboratory The method used for this purpose is called “PCR”:
“Polymerase Chain Reaction”PCR is the amplification of DNA chains using DNA polymerase using a method that has repeating cycles
![Page 51: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/51.jpg)
Squence cDNA
5’ATGGCCCTGTGGATGCGCCTCCTGCCCCTGCTGGCGCTGCTGGCCCTCTGGGGACCTGACCCAGCCGCAGCCTTTGTGAACCAACACCTGTGCGGCTCACACCTGGTGGAAGCTCTCTACCTAGTGTGCGGGGAACGAGGCTTCTTCTACACACCCAAGACCCGCCGGGAGGCAGAGGACCTGCAGGTGGGGCAGGTGGAGCTGGGCGGGGGCCCTGGTGCAGGCAGCCTGCAGCCCTTGGCCCTGGAGGGGTCCCTGCAGAAGCGTGGCATTGTGGAACAATGCTGTACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAACTAG-3’
’Insulin cDNA
5’ATGGCCCTGTGGATGCGC…………………GGAGAACTACTGCAACTAG-3’
![Page 52: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/52.jpg)
DNA polymerase
![Page 53: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/53.jpg)
![Page 54: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/54.jpg)
![Page 55: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/55.jpg)
![Page 56: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/56.jpg)
![Page 57: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/57.jpg)
![Page 58: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/58.jpg)
![Page 59: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/59.jpg)
Route to the Production by Bacteria of Human Insulin
A fermentor used to grow recombinant bacteria.
This is the step when gene cloning takes place.
The single recombinant plasmid replicates within a cell. Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid – and the human insulin gene.
One cell with the recombinant plasmid
![Page 60: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/60.jpg)
Route to the Production by Bacteria of Human Insulin
The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant
human insulin gene.
![Page 61: Kuliah Kloning](https://reader036.vdokumen.com/reader036/viewer/2022062223/577c84bf1a28abe054ba2ba0/html5/thumbnails/61.jpg)
End
See ya ………