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AssigneeUniversity of Medicine and Dentistry of New Jersey
InventorsLi Fang, Weinning Jiang, Masanori Mitta, Masayori Inouye
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A method of inhibiting the translation of bacterial mRNA is disclosed. The method comprises overexpressing in a bacterium an mRNA which contains a sequence which is complementary to the anti-downstream box region of the 16S rRNA. RNA and DNA constructs for the overexpression of the mRNA of the invention are disclosed. Further, there are disclosed isolated DNA constructs that direct the prolonged expression of a heterologous gene in a cold-shocked bacterium at reduced temperature. The construct can comprise a promoter region of a cold-shocked inducible gene. The replication vehicle comprising such DNA constructs and a method for overexpressing a heterologous gene in a bacterium transformed with such a replication vehicle are also disclosed.
A method for inhibiting protein translation in a bacterium comprising overexpressing in the bacterium an mRNA selected from the group consisting of cspA mRNA, cspB mRNA and csdA mRNA, which comprises an initiation codon and a downstream box nucleotide sequence 3′ to the initiation codon which sequence is complementary to the antidownstream box of the 16S rRNA of the bacterium, and allowing the mRNA to anneal to the anti-downstream box, thereby binding the 16S rRNA and inhibiting translation of other bacterial mRNAs.
The method of claim 2 wherein the overexpressing is by transforming the bacterium with a cloning vector containing a DNA sequence which transcribes the mRNA.
A oligonucleotide RNA construct comprising an initiation codon and a downstream box nucleotide sequence 3′ to the initiation codon which sequence is at least in part complementary to the anti-downstream box of the 16S rRNA of a bacterium, wherein the RNA is an isolated RNA construct or is transcribed from an isolated DNA construct and wherein the downstream box which is complementary to at least a part of the 16S rRNA of a bacterium is selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22.
An isolated oligonucleotide DNA construct which transcribes an mRNA comprising an initiation codon and a downstream box nucleotide sequence 3′ to the initiation codon which sequence is complementary to at least a part of the anti-downstream box of the 16S rRNA of a bacterium, and wherein the mRNA transcribed by the DNA comprises mRNA selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22.
The replication cloning vector which contains a DNA promoter sequence operably linked to a DNA sequence which codes for an mRNA which comprises an initiation codon and a downstream box nucleotide sequence 3′ to the initiation codon which sequence is complementary to at least a part of the anti-downstream box of the 16S rRNA of a bacterium and which mRNA comprises a sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22.
A process to modify the cold shock response of a bacterium comprising transforming the bacterium to overproduce part or all of the 5′ untranslated region of mRNA selected from the group of cold shock proteins consisting of cspA, cspB, and csdA and subjecting the bacterium to environmental or growth conditions that elicit the cold shock response of the bacterium.
The method for protein production in a bacterium under environmental or growth conditions that elicit the cold shock response of the bacterium, wherein said method comprises transforming the bacterium with a gene encoding a downstream box, wherein the downstream box is complementary to at least a part of the 16S rRNA of a bacterium is selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22.
An isolated DNA construct that directs the prolonged expression of a heterologous gene in a bacterium under environmental or growth conditions that elicit the cold shock response of the bacterium wherein the DNA construct transcribes mRNA selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:21, and SEQ ID NO:22.
The isolated DNA construct of claim 46 in which the 5′ UTR is that of CspA.
Dependencies are inferred from the claim text.