![]() ![]() In this chapter the various types of host cell will be described first,followed by vector systems and methods for getting DNA into cells.ĢTiga hal yang harus dilakukan untuk mengisolasi gen tertentu dari koleksi urutan DNA rekombinan:molekul rekombinan individu harus secara fisik terpisah satu sama lain,urutan rekombinan harus diperbanyak untuk menyediakan cukup bahan untuk analisis lebih lanjutfragmen target tertentu harus dipilih melalui metode yang didasarkan atas urutan basa spesifik.Kloning gen berkaitan dengan pemilihan dan penggunaan: vektor (molekul pembawa) sel inang (host) di mana vektor dapat diperbanyak.Sel Inang dan Vektor3LO 21: menjelaskan sel inang yang digunakan dalam kloning genThree things have to be done to isolate a gene from a collection of recombinant DNA sequences: the individual recombinant molecules have to be physically separated from each other,the recombinant sequences have to be amplified to provide enough material for further analysis, and the specific fragment of interest has to be selected by some sort of sequence-dependent method. ![]() This is an essential part of most genetic manipulation programmes.Even if the desired result is a transgenic organism, the gene to be used must first be isolated and characterised, and therefore cloning systems are required.Methods for selecting specific sequences are described in Chapter 8.The biology of gene cloning is concerned with the selection and use of a suitable carrier molecule or vector, and a living system or host in which the vector can be propagated. In this chapter I consider the first two of these requirements, which in essence represent the systems and techniques involved in gene cloning. It is a bit like looking for the proverbial needle in a haystack with the added complication that the needle is made of the same material as the haystack! Fortunately the methods available provide a relatively simple way to isolate specific gene sequences.Three things have to be done to isolate a gene from a collection of recombinant DNA sequences: the individual recombinant molecules have to be physically separated from each other,the recombinant sequences have to be amplified to provide enough material for further analysis, and the specific fragment of interest has to be selected by some sort of sequence-dependent method. In some experiments hundreds of thousands of different DNA fragments may be produced, and the isolation of a particular sequence would seem to be an almost impossible task. SEL INANG DAN VEKTOR11Learning Outcome (LO)LO 21: menjelaskan sel inang yang digunakan dalam kloning genLO 22: menjelaskan vektor yang digunakan dalam kloning genLO 23: menjelaskan transformasi LO 24: menjelaskan transfeksiLO 25: menjelaskan metode mikroinjeksi dan biolistik2Host cells and vectors Once recombinant DNA molecules have been constructed in vitro, the desired sequence can be isolated. ![]()
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