{"id":121620,"date":"2022-05-18T14:06:48","date_gmt":"2022-05-18T08:36:48","guid":{"rendered":"https:\/\/www.mapsofindia.com\/my-india\/?p=121620"},"modified":"2022-05-18T14:06:48","modified_gmt":"2022-05-18T08:36:48","slug":"some-restriction-enzymes-break-a-phosphodiester-bond-on-both-the-dna-strands-such-that-only-one-end-of-each-molecule-is-cut-and-these-ends-have-regions-of-single-stranded-dna-bamh1is-one-such-restri","status":"publish","type":"post","link":"https:\/\/www.mapsofindia.com\/my-india\/education\/some-restriction-enzymes-break-a-phosphodiester-bond-on-both-the-dna-strands-such-that-only-one-end-of-each-molecule-is-cut-and-these-ends-have-regions-of-single-stranded-dna-bamh1is-one-such-restri","title":{"rendered":"Some restriction enzymes break a phosphodiester bond on both the DNA strands, such that only one end of each molecule is cut and these ends have regions of single stranded DNA. BamH1is one such restriction enzyme which binds at the recognition sequence, 5\u2019-GGATCC- 3\u2019and cleaves these sequences just after the 5\u2019- guanine on each strand. a. What is the objective of this action? b. Explain how the gene of interest is introduced into a vector. c. You are given the DNA shown below. 5\u2019 ATTTTGAGGATCCGTAATGTCCT 3\u2019 3\u2019 TAAAACTCCTAGGCATTACAGGA 5\u2019 If this DNA was cut with BamHI, how many DNA fragments would you expect? Write the sequence of these double-stranded DNA fragments with their respective polarity. d. A gene M was introduced into E.coli cloning vector PBR322 at BamH1 site. What will be its impact on the recombinant plamids? Give a possible way by which you could differentiate non recombinant to recombinant plasmids."},"content":{"rendered":"<h2>Some restriction enzymes break a phosphodiester bond on both the DNA strands, such that only one end of each molecule is cut and these ends<br \/>\nhave regions of single stranded DNA. BamH1is one such restriction enzyme which binds at the recognition sequence, 5\u2019-GGATCC- 3\u2019and cleaves these sequences just after the 5\u2019- guanine on each strand.<br \/>\na. What is the objective of this action?<br \/>\nb. Explain how the gene of interest is introduced into a vector.<br \/>\nc. You are given the DNA shown below.<br \/>\n5\u2019 ATTTTGAGGATCCGTAATGTCCT 3\u2019<br \/>\n3\u2019 TAAAACTCCTAGGCATTACAGGA 5\u2019<br \/>\nIf this DNA was cut with BamHI, how many DNA fragments would you expect? Write the sequence of these double-stranded DNA fragments with their respective polarity.<br \/>\nd. A gene M was introduced into E.coli cloning vector PBR322 at BamH1 site. What will be its impact on the recombinant plamids? Give a possible way by which you could differentiate non recombinant to recombinant plasmids.<\/h2>\n<h3>Answer.<br \/>\na. The two different DNA molecules will have compatible ends to<br \/>\nrecombine. (\u00bd mark)<br \/>\nb. Restriction enzyme cuts the DNA of the vector and then ligates the<br \/>\ngene of interest into the DNA of the vector.<br \/>\nc. 2 fragments (\u00bd mark)<br \/>\n5\u2019 ATTTTGAG 3\u20195\u2019GATCCGTAATGTCCT 3\u2019<br \/>\n3\u2019 TAAAACTCCTAG 5\u2019.3\u2019GCATTACAGGA 5\u2019<br \/>\nd. BamH1 site will affect tetracycline antibiotic resistance gene,<br \/>\nhence the recombinant plasmids will lose tetracycline resistance<br \/>\ndue to inactivation of the resistance gene.<br \/>\nRecombinants can be selected from non recombinants by plating<br \/>\ninto a medium containing tetracycline, as the recombinants will not<br \/>\ngrow in the medium because the tetracycline resistance gene is<br \/>\ncut.<\/h3>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Some restriction enzymes break a phosphodiester bond on both the DNA strands, such that only one end of each molecule is cut and these ends have regions of single stranded DNA. BamH1is one such restriction enzyme which binds at the recognition sequence, 5\u2019-GGATCC- 3\u2019and cleaves these sequences just after the 5\u2019- guanine on each strand. [&hellip;]<\/p>\n","protected":false},"author":21830,"featured_media":121571,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[7],"tags":[],"class_list":{"0":"post-121620","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-education"},"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/posts\/121620","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/users\/21830"}],"replies":[{"embeddable":true,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/comments?post=121620"}],"version-history":[{"count":1,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/posts\/121620\/revisions"}],"predecessor-version":[{"id":121621,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/posts\/121620\/revisions\/121621"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/media\/121571"}],"wp:attachment":[{"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/media?parent=121620"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/categories?post=121620"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.mapsofindia.com\/my-india\/wp-json\/wp\/v2\/tags?post=121620"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}