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7/30/2019 Arida Fauziyah B1J011713 Class C

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T u g a s T e r s s t r u k t u r B i o l o g i M o l e k u l e r 2 0 1 3 | 1

LAPORAN TUGAS TERSTRUKTUR BIOLOGI MOLEKULER 2013

KELAS C ROMBONGAN 4

1. menjelaskan TEXTBOX di depan dosen setelah diskusi dengan TA paling lambat KAMIS, 11 April 2013,Jam 16.00 WIB

2. menuliskan dan megirimkan penjelasan TEXTBOX setelah menjelaskan di depan dosen [email protected] lambat KAMIS, 18 April 2013, Jam 22.00 WIB.

3. tulis nama file laporan Anda: Kelas, Rombongan, NIM, NamaContoh: C.4. B1J011169 HANI SEPTIANI

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B1J011169 HANI SEPTIANI

LO 42: menjelaskan sel inang yang digunakan dalam kloning gen

TEXTBOX: Gene cloning utilises the characteristics of living systems to propagate recombinant DNAmolecules; in essence this can be considered as a form of molecular agriculture (63.1).

TA:

LAPORAN TUGAS:Tulis di sini

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B1J011171 DWI AGUSTINALO 43: menjelaskan vektor yang digunakan dalam kloning gen

TEXTBOX: Gene cloning is achieved by using a vector (carrier) to propagate the desired sequence in ahost cell. Choosing the right vector/host combination is one of the critical stages of a cloning procedure(63.2).

TA:


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B1J011173 ARIDA FAUZIYAH

LO 44: menjelaskan sel inang prokariotik

TEXTBOX: The bacterium Escherichia coli is the most commonly used prokaryotic host cell, with a widevariety of different strains available for particular Applications (65.1).

TA: Anita Maulinda


Prokaryotic hosts

There are so much prokaryotic hosts available for any specific or not specific vectors,

but we discuss here about an ideal host cell. Three basic requirements that should be fulfil by

an ideal host cell. First, an ideal host cell should be easy to control and develop. Second, it

should be available as wide variety of genetically defined strains, and the third is it shouldaccept a wide range of vectors. And what a wonderful Escherichia coli, its fulfil all of that

requirements. Thats whyE. coli used in many cloning procedure, andE. coli has been studied

in great detail so we can get so much basic genetic information about E. coli for cloning

experiments. Beside that, many different strains of E. coli were isolated. That last fact

embraces the third requirements of an ideal host.

Why E. coli? What is E. coli?Escherichia coli is a negative gram bacterium with a

single chromosome that packed in a compact structure called nucleoid. E. coli has a genome

size about 4.6 x 106

base pairs, and all of complete sequences now are known. As we know that

bacterium is a prokaryotic group, so if we talking about gene synthesis, we found any some

differences between prokaryotic and us as an eukaryotic. There is no post-transcriptional

modification of primary transcript in transcription phase. Eukaryotic cell has intron to splice in

post-transcription modification, but prokaryotic didnt have. E. coli transcripts all of primary

DNA and then translating it. E. coli has been considered as a simplest host cell, much of
mailto:[email protected]:[email protected]:[email protected]


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cloning experiments involves E. coli hosts with many different genetically defined strains for

specific application.

How about the other bacterium?Bacillus,Pseudomonas, Streptomyces, why they cant?

They still can involve in cloning experiments, even some of them have fewer suitable vectors

for use, but still, they can cause a problem. The problem is about getting recombinant DNA

into the cell and direct introduction of ligated recombinant DNA into the host cell. Meanwhile,

this type of application needs an efficient and reliable procedure to maximise the yield of

cloned fragments. And again, E. coli more sensible to isolates the requirements sequence and

then introduce the purified DNA into the target host. This approach can covered many

disadvantages of using other bacterium beside E. coli. This is gonna be greater when vectors

can function in target host and in E. coli as a shuttle vector.

Inang prokariotik

Tersedia banyak sel inang prokariotik untuk beberapa vektor yang spesifik maupun non

spesifik. Dari sekian banyak inang prokaritok tersebut, terdapat sel prokariotik yang ideal untuk

dijadikan inang. Tiga hal penting yang harus dipenuhi untuk menjadi sel inang yang ideal.

Pertama, sel inang yang ideal harus mudah dikontrol dan berkembang, kedua sel inang ideal

harus memiliki banyak varietas strain yang diketahui agar dapat menerima variasi dari

kelompok-kelompok vektor. Escherichia coli lah yang memenuhi ketiga kriteria utama

tersebut. Faktanya, E. coli sering digunakan dalam banyak eksperimen mengenai kloning, hal

ini diperkuat karena E. coli telah diteliti pada tingaktan yang lebih detail sehingga kita

mendapatkan informasi genetik dasar yang sangat membantu dalam proses eksperimen. Selain

itu, sudah banyak sekali sekuens yang berbeda dariE. coli yang telah diisolasi.

E. coli adalah bakteri gram negatif dengan kromosom tunggal yang tersusun dalam

sebuah struktur yang disebut dengan nucleoid. Ukuran genom dari E. coli adalah 4.6 x 106

pasang basa, dan kesemua sekuens lengkap tersebut sekarang telah diketahui. Berbeda dengan

sel eukariotik sel proakariotik tidak mengalami modifikasi setelah transkripsi pada sintesis

DNA. Sel eukariotik akan melakukan intron-splicing pada modifikasi pasca transkripsi, sel

prokariotik tidak memilki ekson maupun intron untuk di-splicing, sehingga sel prokaritik

mentranskrip semua primer transkrip dan kemudian mentranslasikannya. E. coli

dipertimbangkan sebagai sel inag yang paling sederhana dengan keberadaan strain-strain yang

berbeda untuk penerapan yang spesifik.

Bakteri selainE. coli, sepertiBacillus,Pseudomonas, dan Streptomyces dapat dilibatkan

dalam percobaan-percobaan kloning, bahkan ada beberapa diantaranya yang memilki strain

spesifik untuk beberapa vektor yang spesifik pula. Namun, bakteri-bakteri ini masih beresiko

menimbulkan masalah, diantaranya pada saat pengambilan DNA rekombinan pada sel dan

introduksi langsung DNA rekombinan yang mengalami ligase pada sel inang. Padahal, tipe

penerapan ini membutuhkan prosedur yang mudah dan efisien untuk memaksimalkan hasil dari


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fragmen-fragmen yang terkloning. Dalam hal ini, E.coli lebih mampu untuk mendapakatkan

isolasi DNA yang dibutuhkan yang nantinya akan diintroduksi berupa DNA yang telah

dimurnikan kepada sel target. Pendekatan ini dapat menutupi kekurangan-kekurangan dari

penggunaan bakteri sebagai sel inang selain selE. coli. Hal ini akan menjadi lebih baik apabila

vektor tersebut dapat berfungsi pada sel inang target dan selE. coli sebagai sel inang pemindah

(shuttle vector).

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B1J011175 R.A. NUR ADITA K.S.

LO 45: menjelaskan sel inang eukariotik

TEXTBOX: Microbes (such as yeast) and mammalian cell lines are two examples of eukaryotic hostcells that have become widely used in gene manipulation (66.1).

TA:


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B1J007125 TRININGSIH

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel iang E. coli

TEXTBOX: Plasmids are extrachromosomal genetic elements that are not essential for bacteria tosurvive but often confer advantageous traits (such as antibiotic resistance) on the host cell (66.2).

TA:


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B1J008143 WONDO PRIYANTO

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel iang E. coli

TEXTBOX: pBR322 is a very famous plasmid and has all the essential requirements for a cloningvector relatively small size, useful restriction enzyme sites, an origin of replication, and antibioticresistance genes (68.1).

TA:


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B1J009016 DIODORA ADEA AGINTHA

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel inang E. coli

TEXTBOX: Multiple cloning sites (polylinkers) increase the flexibility of vectors by providing a range ofrestriction sites for cloning (69.1).

TA:LAPORAN TUGAS:Tulis di sini

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B1J009048 DWI NURCAHYO A

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel inang E. coli

TEXTBOX: Plasmid vectors have an upper size limit for efficient cloning, which can sometimes restricttheir use where a large number of clones is required. In this case it makes sense to clone longer DNAfragments, and a different vector system is needed (69.2).

TA:


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B1J009090 SUKENDA

LO 47: menjelaskan vektor bakteriofaga yang digunakan dalam sel inang E. coli

TEXTBOX: Bacteriophages are essentially bacterial viruses and usually consist of a DNA genomeenclosed in a protein head (capsid). As with other viruses, they depend on the host cell for theirpropagation and do not exist as free-living organisms (71.1)

TA:



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B1J009124 RAHMAT HIDAYAT

LO 48: menjelaskan vektor bakteriofagayang digunakan dalam sel inang E. coli

TEXTBOX: Bacteriophage has played a major role in the development of bacterial genetics and

molecular biology. In addition to fundamental aspects of gene regulation, has been used as the basisfor a wide variety of cloning vectors (72.1).

TA:


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B1J009126 KARTIKA FEBRIANA

LO 49: menjelaskan vektor yang digunakan dalam sel inang eukariotik

TEXTBOX: A range of plasmid-based vectors for the yeast Saccharomyces cerevisiae was developed

from the naturally occurring yeast 2m plasmid (81.1).

TA:


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B1J009130 FALAH FARIKHATIN

LO 49: menjelaskan vektor yang digunakan dalam sel inang eukariotik

TEXTBOX: Vectors for use in plant and animal cells have properties that enable them to function inthese cell types; they are often more specialised than the basic primary cloning vectors such

as (82.1).

TA:


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B1J009162 ELIARIZKI UTAMI

LO 50: menjelaskan kromosom artifisialTEXTBOX: Artificial chromosomes are elegantly simple vectors that mimic the natural constructionof chromosomal DNA, with telomeres, a centromere, and an origin of replication in addition to featuresdesigned for ease of use, such as selectable markers (83.1).

TA:


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B1J009163 NURHADI EKO F.

LO 51: menjelaskan transformasi

TEXTBOX: Transformation ofE. colicells with recombinant plasmid DNA is one of the classic

techniques of gene manipulation (84.1).TA:


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B1J009180 TOCHIRUN

LO 51: menjelaskan transformasi

TEXTBOX: Transformation efficiency is often a limiting factor in using the technique, and this may becritical if the aim of the procedure is to prepare a representative clone bank (85.1).

TA:


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B1J010094 NUGROHO YOGA PRANA

LO 52: menjelaskan transfeksi

TEXTBOX: Packaging recombinant bacteriophage DNA in vitro mimics the normal process that occursduring phage maturation and assembly and has proved to be a very useful method for the constructionof genomic libraries (86.1).

TA:



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B1J010171 NOOR WAHYU ANISAH

LO 53: mejelaskan metode mikroinjeksi dan biolistik

TEXTBOX: Introducing recombinant DNA into eukaryotic cells can involve biological methods or one ofa range of techniques such as electroporation, microinjection, or biolistics (86.2)

TA:


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