ʻO Gene cloning ka hana o ke kopeʻana i nā kope, aiʻole nā olonā, o kahi'āpana hoʻokahi. Keʻikeʻia kekahiʻano, hiki ke hoʻohanaʻia nā molonika ma nāʻano he nui o nāʻimi noiʻi me nāʻoihanaʻoihana. ʻO ke'enekeneke'enewai keʻano o ke kinouʻana i nā gen i loko o nā meaola hou a iʻole e hoʻololi i ke kaʻina DNA e hoʻololi i ka huahana waiwai. Hoʻoponopono ka hanaʻenehana waiwai ma muli o ka hiki iā mākou ke hana i nā kaʻina hana.
01 Hoʻonākiuki Hua'ōlelo
02 Mau Mau Enzymes
ʻO ka loaʻaʻana o nā enzymes i kapaʻia he endonucleases hoʻolālā he mea nui ia i ka hoʻolele pūmua . Hoʻopiʻi kēia mau enzymes i ka DNA ma nā wahi kiko'ī e like me ka nucleotide. Hiki ke hoʻokaʻawaleʻia he mau haneli haneli likeʻole, hiki ke hana i ka DNA ma kahi kahua kaʻawale, mai nāʻanoʻeha o nā mea maʻi. ʻO ka DNA i kālaiʻia i kahi enzyme hoʻonāuki e loaʻa mai i nā'āpana liʻiliʻi he nui, heʻano likeʻole. Hiki ke hoʻokaʻawaleʻia kēia mau mea me ka electrophoresis gel a chromatography.
03 Ke kohopili
ʻO ka hoʻomaʻemaʻeʻana i ka DNA mai kahi moʻomeheu kamela, a iʻole eʻoki ana iā ia me ka hoʻohanaʻana i nā enzymes palenaʻole,ʻaʻole ia he mea nui ināʻaʻole hiki iā mākou ke nānā i ka DNA -ʻo ia hoʻi, e huli i kahi e nānā ai inā i loaʻa i kāu huapalapala kekahi mea, ʻokiʻia i loko. ʻO hoʻokahi o ka hana e hana ai kēia ma ka gel electrophoresis. Hoʻohanaʻia nā Gels no nā kumu likeʻole, mai ka nānāʻana i ka DNA i kāʻiliʻia no kaʻikeʻana i nā inserts DNA a me nā kīkēkē.
04 Hui pū me DNAʻelua
I ka huliʻana o ka genetic, pono pinepine e pili iʻelua mauʻokoʻa o DNA, e hana i kahi kaula hou, aiʻole e hoʻopau i kahi kapo iʻokiʻia me nā hoʻolālā kūpilikiʻi. Hiki i nā enzymes i kapaʻiaʻo DNA ligases ke hana i nā pilina ma waena o nā kaula nucleotide. ʻO nā enzymesʻo DNA polymerase I a me polynucleotide kinase he mea nui nō hoʻi i kēia kaʻina hana, no ka hoʻopihaʻana i nāʻawa a iʻole phosphorylating i nā pae '5', i kēlā me kēia.
05 E kohoʻana i ka DNA pūnaewele pono'ī
ʻO nā'āpana o ka DNA liʻiliʻiʻaʻole i pili i ka genome bacteria, akā hiki iā lākou ke hana pilikino pono'ī, i kapaʻia he plasmids. Hoʻohanaʻia nā plasmids e like me nā vera e lawe i nā genes ma waena o nā meaʻano. I loko o ka biotechnology, i ka manawa i hoʻonuiʻia ai ka lili huahua a uaʻokiʻokiʻia ka gen a me ka plasmid e nā palena enzymes, ua hoʻohuiʻia lākou me ka hanaʻana i ka mea iʻikeʻia he DNA hou. Hiki ke hoʻohanaʻia ka DNA (bacteriophage) DNA e like me ke kikowaena, e like me nā mea maʻamau, nā plasmids recombinant i loko o nā genes bacteriophage.
ʻOihana 06 no ka Move i kahi Vector i loko o kahi Pūnaewele Pūnaewele
ʻO ke kaʻina hana o ka hoʻoiliʻana i nā meaʻokoʻa ma luna o kahi kiʻi e like me ka plasmid, i loko o nā pūnaewele hou, ua kapaʻiaʻo ka loli. Pono kēiaʻano hana eʻikeʻia nā pūnaewele pūnaewele i kahi hoʻololi o ka'eneka i mea e "maʻalahi" aiʻole no kekahi manawa kūpono i ka veʻa. ʻO ka koho ponoʻana kahi hoʻokahiʻanoʻano. ʻO ka nui nui o ka plasmid,ʻo ka haʻahaʻa ka mea maikaʻi i laweʻia e nā pūnaewele. Ua maʻalahi ka maʻalahi o nā'āpana DNA ma ka hoʻohanaʻana i ka bacteriophage, reprovirus a me nā vectors viral'ē aʻe aiʻole ka mea hana i loko o keʻano i kapaʻia he transduction. Hoʻohana pinepineʻia nā lāmākū a me nā veca viral i loko o ka lāʻau lapaʻau hou akā hiki ke hoʻokomo i ka DNA ma nā'āpana o kā mākou chromosoma i kahiʻole mākou e makemake ai, e loaʻa ai nā pilikia a me ka maʻi kanesa.
07 Nā Hanana e koho i nā Organic Transgenic
ʻAʻole nā pūnaewele a pau e lawe i ka DNA i ka manawa e hoʻololiʻia ai. He mea nui e loaʻa i kahi ala eʻike ai i nā mea hana. ʻO ka mea maʻamau, hiki ke kohoʻia nā'ōmene i nā genes no ke kū'ēʻana i ka lāʻau a me nā cell transgenic ma muli o ka hōʻikeʻana o kēlā mau genes a me ko lākou hiki ke ulu ma luna o kaʻili i loko o kēlā lāʻauʻae. Hiki ke kohoʻia kekahi mauʻano'ē aʻe o ka kohoʻana ma kaʻike o nā mea'ē aʻe e like me ka pūnaehana x-gal / lacZ , aiʻole ka protein protein fluorescence, eʻae i ka kohoʻana ma muli o ke kala a me ka fluorescence.