TugasTersstruktur Biologi Molekuler 2013 |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 ke hendropram@facebook.com paling lambat KAMIS, 18 April 2013, Jam 22.00 WIB.

3.

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

LO 42: menjelaskan sel inang yang digunakan dalam kloning gen

40 TEXTBOX: Gene cloning utilises the characteristics of living systems to propagate recombinant DNA molecules; in essence this can be considered as a form of molecular agriculture (63.1). TA: LAPORAN TUGAS: Tulis di sini

B1J011171 DWI AGUSTINA

LO 43: menjelaskan vektor yang digunakan dalam kloning gen

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

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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 wide variety of different strains available for particular Applications (65.1). TA: Anita Maulinda LAPORAN TUGAS: Tulis di sini

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 should accept a wide range of vectors. And what a wonderful Escherichia coli, its fulfil all of that
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requirements. Thats why E. coli used in many cloning procedure, and E. 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

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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 dari E. 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 selain E. coli, seperti Bacillus, 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 sel E. coli. Hal ini akan menjadi lebih baik apabila vektor tersebut dapat berfungsi pada sel inang target dan sel E. coli sebagai sel inang pemindah ( shuttle vector ).

B1J011175 R.A. NUR ADITA K.S.

LO 45: menjelaskan sel inang eukariotik

43 TEXTBOX: Microbes (such as yeast) and mammalian cell lines are two examples of eukaryotic host cells that have become widely used in gene manipulation (66.1). TA: LAPORAN TUGAS: Tulis di sini

B1J007125 TRININGSIH

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel iang E. coli
44 TEXTBOX: Plasmids are extrachromosomal genetic elements that are not essential for bacteria to survive but often confer advantageous traits (such as antibiotic resistance) on the host cell (66.2). TA: LAPORAN TUGAS: Tulis di sini

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 cloning vector relatively small size, useful restriction enzyme sites, an origin of replication, and antibiotic resistance genes (68.1). TA: LAPORAN TUGAS: Tulis di sini

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B1J009016 DIODORA ADEA AGINTHA

LO 46: menjelaskan vektor plasmid yang digunakan dalam sel inang E. coli
46 TEXTBOX: Multiple cloning sites (polylinkers) increase the flexibility of vectors by providing a range of restriction sites for cloning (69.1). TA: LAPORAN TUGAS: Tulis di sini

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 restrict their use where a large number of clones is required. In this case it makes sense to clone longer DNA fragments, and a different vector system is needed (69.2). TA: LAPORAN TUGAS: Tulis di sini

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B1J009090 SUKENDA

LO 47: menjelaskan vektor bakteriofaga yang digunakan dalam sel inang E. coli
48 TEXTBOX: Bacteriophages are essentially bacterial viruses and usually consist of a DNA genome enclosed in a protein head (capsid). As with other viruses, they depend on the host cell for their propagation and do not exist as free-living organisms (71.1) TA: LAPORAN TUGAS: Tulis di sini

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B1J009124 RAHMAT HIDAYAT

LO 48: menjelaskan vektor bakteriofaga yang 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 basis for a wide variety of cloning vectors (72.1). TA: LAPORAN TUGAS: Tulis di sini

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B1J009126 KARTIKA FEBRIANA

LO 49: menjelaskan vektor yang digunakan dalam sel inang eukariotik

50 TEXTBOX: A range of plasmid-based vectors for the yeast Saccharomyces cerevisiae was developed from the naturally occurring yeast 2m plasmid (81.1). TA: LAPORAN TUGAS: Tulis di sini

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 in these cell types; they are often more specialised than the basic primary cloning vectors such as (82.1). TA: LAPORAN TUGAS: Tulis di sini

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B1J009162 ELIARIZKI UTAMI

LO 50: menjelaskan kromosom artifisial

TEXTBOX: Artificial chromosomes are elegantly simple vectors that mimic the natural construction of chromosomal DNA, with telomeres, a centromere, and an origin of replication in addition to features designed for ease of use, such as selectable markers (83.1). TA: LAPORAN TUGAS: Tulis di sini

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B1J009163 NURHADI EKO F.

LO 51: menjelaskan transformasi

53 TEXTBOX: Transformation of E. coli cells with recombinant plasmid DNA is one of the classic techniques of gene manipulation (84.1). TA: LAPORAN TUGAS: Tulis di sini

B1J009180 TOCHIRUN

LO 51: menjelaskan transformasi

54 TEXTBOX: Transformation efficiency is often a limiting factor in using the technique, and this may be critical if the aim of the procedure is to prepare a representative clone bank (85.1). TA: LAPORAN TUGAS: Tulis di sini

B1J010094 NUGROHO YOGA PRANA

LO 52: menjelaskan transfeksi

55 TEXTBOX: Packaging recombinant bacteriophage DNA in vitro mimics the normal process that occurs during phage maturation and assembly and has proved to be a very useful method for the construction of genomic libraries (86.1). TA: LAPORAN TUGAS: Tulis di sini

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B1J010171 NOOR WAHYU ANISAH

LO 53: mejelaskan metode mikroinjeksi dan biolistik

56 TEXTBOX: Introducing recombinant DNA into eukaryotic cells can involve biological methods or one of a range of techniques such as electroporation, microinjection, or biolistics (86.2) TA: LAPORAN TUGAS: Tulis di sini